WO2022228495A1 - Preparation method for antibody-drug conjugate, and application - Google Patents

Preparation method for antibody-drug conjugate, and application Download PDF

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Publication number
WO2022228495A1
WO2022228495A1 PCT/CN2022/089726 CN2022089726W WO2022228495A1 WO 2022228495 A1 WO2022228495 A1 WO 2022228495A1 CN 2022089726 W CN2022089726 W CN 2022089726W WO 2022228495 A1 WO2022228495 A1 WO 2022228495A1
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optionally substituted
compound
group
cancer
tautomer
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PCT/CN2022/089726
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French (fr)
Chinese (zh)
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张惠
孟逊
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上海汇连生物医药有限公司
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Priority to CA3173902A priority Critical patent/CA3173902A1/en
Priority to CN202280030781.1A priority patent/CN117337195A/en
Priority to EP22794965.8A priority patent/EP4331613A1/en
Priority to KR1020237041142A priority patent/KR20240006029A/en
Publication of WO2022228495A1 publication Critical patent/WO2022228495A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6889—Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A61K47/68037—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a camptothecin [CPT] or derivatives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic

Definitions

  • the present application relates to the field of biomedicine, in particular to a preparation method and application of an antibody conjugated drug.
  • ADC Antibody Drug Conjugate
  • Cytotoxics are essential for ADC drugs to play a role. Camptothecin drugs have great application prospects.
  • the marketed ADC drugs Trodelvy and Enhertu use camptothecin drugs SN38 and DX-8951f as warhead molecules respectively.
  • camptothecins finally prepared by camptothecins have greatly changed the properties of monoclonal antibodies, and the stability and half-life have been greatly attenuated.
  • the objective response rate (ORR) confirmed by Enhertu reached 79.7%
  • the objective response rate (ORR) confirmed by Enhertu was 37.0 for HER2-low expressing breast cancer %
  • the treatment efficiency was significantly lower than that of breast cancer with high or medium expression.
  • ADC drugs have new symptoms of toxic side effects, such as pneumonia and interstitial lung disease with the drug Enhertu.
  • WO2020233174A1 discloses a class of camptothecin compounds containing a valine-citrulline (Val-Cit)-PAB linker, however, this molecule cannot be coupled with an antibody to obtain a qualified ADC molecule (in qualified ADC products).
  • the aggregate content needs to be less than 5%). Therefore, there is an urgent need in the art to provide more suitable antibody-drug conjugates based on camptothecins (such as ixatecan, belotecan, etc.) to achieve efficient, simple and practical chemical preparation and conjugation, and Improve the pharmaceutical properties, metabolic properties, efficacy and safety of the existing antibody-conjugated drugs, such as improving the stability of ADC molecules, improving the therapeutic window, etc.
  • the present application provides an antibody conjugated drug, an intermediate thereof, a preparation method and an application thereof.
  • the antibody conjugated drug of the present application can realize the wide application of cytotoxic drugs in the field of ADC to treat tumor diseases.
  • the main technical effect of the present application is that the provided novel linker can combine highly hydrophobic anti-tumor drugs, such as topoisomerase inhibitors isanotecan, belotecan, etc., with antibodies through specific chemical methods. Conjugation, the obtained conjugate has high hydrophilicity and stability.
  • the antibody conjugate provided by the application is not easy to produce aggregates when the drug load is higher;
  • the toxin molecules released in the present application are the prototype molecules of topoisomerase inhibitor ixatecan and belotecan. Tests show that the drug molecules have better properties than the drug derivatives released by similar ADC drugs. Good biological activity, safety and other drug-related properties. Therefore, the present application can improve the half-life of the drug in vivo and the concentration of the drug in the tumor tissue, thereby referring to the anti-tumor activity of the drug and/or improving the overall drug treatment window.
  • the application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or A pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the compound comprises the structure represented by the formula (C-Trop-2):
  • L 1a is selected from the group consisting of optionally substituted alkylene, optionally substituted polyethylene glycol, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted alicyclic groups, optionally substituted alicyclic heterocyclyl groups, optionally substituted arylene groups, and optionally substituted heteroarylene groups;
  • L2 comprises optionally substituted polypeptide residues
  • L 3 comprises an optionally substituted spacer group
  • L 2 and/or L 3 comprise optionally substituted polysarcosine residues
  • T contains the drug unit
  • Ab is a ligand capable of binding Trop-2, and m is a number from 1 to 8.
  • the application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a mixture thereof
  • a pharmaceutically acceptable salt, prodrug or solvate
  • the compound comprises a structure selected from the group consisting of:
  • Ab is a ligand capable of binding Trop-2, and m is a number from 1 to 8.
  • the application provides a pharmaceutical composition, which contains the compound described in any one of the application, or its tautomer, meso, racemate, enantiomer, A diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, and optionally a pharmaceutically acceptable carrier.
  • the present application provides a compound containing any one of the present application, or its tautomer, meso, racemate, enantiomer, diastereomer, Use of the mixture form thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, and/or the pharmaceutical composition described in this application in the preparation of a medicament for treating and/or preventing tumors.
  • the present application provides an antibody-drug conjugate, which is formed by coupling the compound of the present application with an antibody.
  • the conjugates of the present application are covalently attached to one or more drug components.
  • the antibody and drug of the present application are coupled covalently (eg, by covalent attachment to a linker, respectively).
  • the present application provides a method for preparing an antibody-drug conjugate, wherein a pair of cysteine residues is generated by reduction of the disulfide chain in the hinge region of the antibody or antibody fragment, and the cysteine
  • the sulfhydryl group in the acid residue undergoes a substitution reaction with the connecting group of the compound in the present application, such as maleimide group, and then the compound of the present application is connected to the cysteine sulfhydryl group of the antibody or antibody fragment to obtain an antibody -Drug conjugate
  • the drug-antibody conjugation rate DAR (for example, m in this application) is controlled according to the reaction conditions, for example, it is usually between 2 and 8.
  • m represents the molar ratio of cytotoxic drug molecules to Ab (also known as DAR, that is, drug-antibody coupling ratio), and m can be an integer or a decimal number, which can be understood as: a single monoclonal antibody molecule and cell
  • the average molar ratio of drug molecules to monoclonal antibody molecules in the antibody-drug conjugate obtained after toxic drug conjugation can generally be determined by hydrophobic chromatography (Hydrophobic-Interaction Chromatography, HIC), reversed-phase high-performance liquid chromatography (Reverse high-performance liquid chromatography) phase HPLC, RP-HPLC), polyacrylamide-SDS gel electrophoresis (SDS PAGE, electrophoresis), liquid chromatograph-mass spectrometer (LC-MS), ultraviolet/visible spectroscopy (UV/Vis), etc. measured.
  • the application provides a method for preparing the compound of the application, comprising the steps of:
  • an amino acid active ester with an amino protecting group N1 with an amino acid to obtain an intermediate M1; in the presence of a condensing agent such as EEDQ (2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline)
  • the intermediate M1 is contacted with substituted/unsubstituted p-aminobenzyl alcohol to obtain the intermediate M2; the N1 of the intermediate M2 is removed to obtain the intermediate M3; contacting a compound of an imide group to obtain an intermediate M4; contacting the intermediate M4 with bis(4-nitrophenyl) carbonate to obtain an intermediate M5; contacting the intermediate M5 with a drug unit .
  • the N1 comprises fluorenylmethoxycarbonyl.
  • the intermediate comprising the amino protecting group N2 is contacted with trifluoroacetic acid, followed by contact with acetyl polysarcosine contacts.
  • the N2 comprises tert-butoxycarbonyl.
  • the application provides a method of preparing a compound of the application comprising the steps of contacting the ligand with the compound of the application under conditions suitable for forming a bond between the ligand and the compound.
  • the ligand is contacted with the compound of the present application in a mixture of buffer and organic solvent.
  • the ligand is contacted with a compound of the present application at about 0 to about 37°C.
  • the following step is included before contacting the ligand with the compound of the present application: reacting the ligand with a reducing agent in a buffer to obtain the reduced ligand.
  • the step of removing the reducing agent is included.
  • the removing the reducing agent comprises subjecting the reaction product to a desalting column and/or ultrafiltration.
  • the reducing agent is selected from the group consisting of tris(2-carboxyethyl)phosphine hydrochloride (TCEP), beta-mercaptoethanol, beta-mercaptoethylamine hydrochloride, and dithiothreose Alcohol (DTT).
  • TCEP tris(2-carboxyethyl)phosphine hydrochloride
  • beta-mercaptoethanol beta-mercaptoethylamine hydrochloride
  • DTT dithiothreose Alcohol
  • the buffer is selected from the group consisting of potassium dihydrogen phosphate-sodium hydroxide (KH 2 PO 4 -NaOH)/sodium chloride (NaCl)/diethyltriaminepentaacetic acid (DTPA) Buffer, disodium hydrogen phosphate-citric acid/sodium chloride (NaCl)/diethyltriaminepentaacetic acid (DTPA), boric acid-borax/sodium chloride (NaCl)/diethyltriaminepentaacetic acid (DTPA) , histidine-sodium hydroxide/sodium chloride (NaCl)/diethyltriaminepentaacetic acid (DTPA) and PBS/diethyltriaminepentaacetic acid (DTPA).
  • KH 2 PO 4 -NaOH potassium dihydrogen phosphate-sodium hydroxide
  • DTPA diethyltriaminepentaacetic acid
  • Buffer dis
  • the organic solvent is selected from the group consisting of acetonitrile (ACN), dimethylformamide (DMF), dimethylacetamide (DMA) and dimethylsulfoxide (DMSO).
  • ACN acetonitrile
  • DMF dimethylformamide
  • DMA dimethylacetamide
  • DMSO dimethylsulfoxide
  • the volume ratio of the organic solvent in the buffer solution and the organic solvent mixture does not exceed 30%.
  • This application finds that the antibody-drug conjugates in this application, compared with traditional antibody-drug conjugates, due to the large hydrophobicity of ixatecan and belotecan drugs, the introduction of polysarcosine can The hydrophilicity of the conjugate is greatly increased, thereby making the obtained antibody-drug conjugate more stable and less prone to polymerization as a whole.
  • the present application provides carbamate, which can undergo rapid 1,6-elimination after enzymatic cleavage to release the drug, and has better stability and biological activity in vitro and in vivo. Based on the above findings, the present application has been completed.
  • Figure 1 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 1 of the present application
  • Figure 2 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 2 of the present application
  • Figure 3 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 2 of the present application
  • Figure 4 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 3 of the present application
  • Figure 5 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 4 of the present application
  • Figure 6 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 4 of the present application
  • Figure 7 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 5 of the present application
  • Figure 8 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 5 of the present application
  • Figure 9 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 6 of the present application.
  • Figure 10 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 6 of the present application
  • Figure 11 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 7 of the present application
  • Figure 12 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 7 of the present application
  • Figure 13 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 8 of the present application
  • Figure 14 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 8 of the present application
  • Figure 15 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 9 of the present application
  • Figure 16 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 9 of the present application
  • Figure 17 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 10 of the present application
  • Figure 18 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 10 of the present application
  • Figure 19 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 11 of the present application
  • Figure 20 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 11 of the present application
  • Figure 21 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 12 of the present application
  • Figure 22 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 12 of the present application
  • Figure 23 shows the concentration change diagram of the accelerated stability experiment of some antibody conjugates of the present application.
  • Figure 24 shows a graph showing the increase of aggregates in the accelerated stability experiment of some antibody conjugates of the present application.
  • Figure 25 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 13 of the present application
  • Figure 26 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 14 of the present application
  • Figure 27 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 15 of the present application
  • Figure 28 shows a comparison chart of the efficacy results of some antibody conjugates of the present application in human lung squamous cell carcinoma NCI-H2170;
  • Figure 29 is a graph showing the changes in the body weight of mice in the in vivo efficacy experiment of partial antibody conjugates of the present application in human lung squamous cell carcinoma cells NCI-H2170.
  • the term "ligand” generally refers to a macromolecular compound capable of recognizing and binding to an antigen or receptor associated with a target cell.
  • the role of the ligand can be to present the drug to the target cell population that binds to the ligand, including but not limited to protein hormones, lectins, growth factors, antibodies, or others that can bind to cells, receptors and/or antigens molecule.
  • the ligand can be represented as Ab, and the ligand antigen forms a bond with the connecting unit through the heteroatom on the ligand, which can be an antibody or an antigen-binding fragment thereof, and the antibody can be selected from chimeric antibodies, human-derived antibody, fully human, or murine; the antibody may be a monoclonal antibody.
  • the antibody may be an antibody or antigen-binding fragment thereof targeting a target selected from the group consisting of: HER2.
  • alkyl generally refers to a residue derived from an alkane by removal of a hydrogen atom. Alkyl groups can be substituted or unsubstituted, substituted or unsubstituted.
  • alkyl generally refers to a saturated straight-chain or branched aliphatic hydrocarbon group having a residue derived by removing a hydrogen atom from the same carbon atom or two different carbon atoms of the parent alkane, which may be a group containing 1 to A straight or branched chain group of 20 carbon atoms, eg 1 to 12 carbon atoms, eg, an alkane alkyl group containing 1 to 6 carbon atoms.
  • alkyl groups include, but are not limited to, methyl, ethyl, propyl, propyl, butyl, and the like.
  • Alkyl groups may be substituted or unsubstituted, substituted or non-substituted, for example when substituted, substituents may be substituted at any available point of attachment, and the substituents may be independently optionally selected from alkyl groups , alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy , heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and one or more substituents in oxo, such as hydrogen, protium, deuterium, tritium, halogen, -NO 2 , - CN, -OH,
  • alkylene generally refers to a saturated straight or branched chain aliphatic hydrocarbon group having 2 hydrogen atoms derived from the same or two different carbon atoms of the parent alkane by removing two hydrogen atoms.
  • residue which may be a straight or branched chain group containing 1 to 20 carbon atoms, for example, the term “methylene” may refer to a residue derived from a group of 1 carbon atom by removing two hydrogen atoms base.
  • a methylene group may be substituted or unsubstituted, substituted or non-substituted; eg, an alkylene group containing from 1 to 12 carbon atoms, eg, an alkylene group containing from 1 to 6 carbon atoms.
  • Non-limiting examples of alkylene groups include, but are not limited to, methylene ( -CH2- ), 1,1-ethylene (-CH( CH3 )-), 1,2-ethylene ( -CH2) CH 2 )-, 1,1-propylene (-CH(CH 2 CH 3 )-), 1,2-propylene (-CH 2 CH(CH 3 )-), 1,3-propylene ( -CH2CH2CH2- ), 1,4 - Butylene ( -CH2CH2CH2CH2- ) and 1,5 - Butylene ( -CH2CH2CH2CH2CH2- ) Wait.
  • Alkylene may be substituted or unsubstituted, substituted or non-substituted, for example, when substituted, substituents may be substituted at any available point of attachment, which may be independently optionally selected from alkanes group, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy substituted by one or more substituents in the group, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo, such as hydrogen, protium, deuterium, tritium, halogen, -NO 2 , -CN, -OH, -SH, -NH 2 , -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH
  • alkenyl generally refers to a straight or branched chain hydrocarbon group containing one or more double bonds.
  • alkenyl groups include allyl, homoallyl, vinyl, crotyl, butenyl, pentenyl, hexenyl, and the like.
  • C2-6 alkenyl groups having more than one double bond include butadienyl, pentadienyl, hexadienyl, and hexatrienyl and branched forms thereof.
  • the position of the unsaturated bond (double bond) can be anywhere in the carbon chain.
  • Alkenyl groups can be substituted or unsubstituted.
  • alkenylene generally refers to residues having two hydrogen atoms removed from the carbon atoms of an olefin.
  • alkenylene groups can be substituted or unsubstituted.
  • alkynyl generally refers to unsaturated straight or branched chain alkynyl groups such as ethynyl, 1-propynyl, propargyl, butynyl, and the like. Alkynyl groups can be substituted or unsubstituted.
  • alkynylene generally refers to residues having two hydrogen atoms removed from the carbon atoms of an alkyne.
  • it can be ethynylene, propynylene, propargylene, butynylene and the like.
  • Alkynylene groups can be substituted or unsubstituted.
  • aryl generally refers to residues having an aromatic ring derived by removing one hydrogen atom.
  • aromatic ring may refer to a 6- to 14-membered all-carbon monocyclic or fused polycyclic ring (ie, rings that share adjacent pairs of carbon atoms) having a conjugated pi-electron system, and may be 6 to 10 membered, such as benzene and Naphthalene.
  • the aromatic ring can be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring linked to the parent structure is an aryl ring.
  • Aryl may be substituted or unsubstituted, and when substituted, the substituent may be one or more of the following groups independently selected from the group consisting of: alkyl, alkenyl, alkynyl, alkoxy, alkane Thio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio , and heterocycloalkylthio.
  • Aryl groups can be substituted or unsubstituted.
  • arylene generally refers to a residue having two hydrogen atoms removed from a carbon atom of an aromatic ring.
  • phenylene and naphthylene may be mentioned.
  • Arylene groups can be substituted or unsubstituted.
  • heteroaryl generally refers to a residue having a hydrogen atom removed from a carbon atom of a heteroaromatic ring.
  • heteromatic ring refers to a heteroaromatic system comprising 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms may be selected from the group consisting of oxygen, sulfur and nitrogen.
  • Heteroaryl can be 5 to 10 membered, 5 membered or 6 membered, such as furanyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazole Base et al.
  • the heteroaryl ring can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is a heteroaryl ring.
  • Heteroaryl groups can be optionally substituted or unsubstituted, and when substituted, the substituents can be one or more of the following groups independently selected from the group consisting of: alkyl, alkenyl, alkynyl, alkoxy group, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, ring Alkylthio, and heterocycloalkylthio. Heteroaryl groups can be substituted or unsubstituted.
  • heteroarylene generally refers to a residue having two hydrogen atoms removed from a carbon atom of a heteroaromatic ring.
  • it may be a furanylene group, a thienylene group, a pyridylene group, a pyrrolidine group, a pyrimidinylene group, a pyrazinylene group, an imidazolylylene group, a tetrazolium group, and the like.
  • a heteroarylene group can be substituted or unsubstituted.
  • alicyclic group generally refers to a residue having a hydrogen atom removed from the same carbon atom or a plurality of different carbon atoms of an alicyclic ring.
  • cycloalkane generally refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon, the carbocyclic ring containing 3 to 20 carbon atoms, may contain 3 to 12 carbon atoms, may contain 3 to 10 carbon atoms, may Contains 3 to 8 carbon atoms.
  • Non-limiting examples of alicyclic groups include cyclopropanyl, cyclobutanyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclopentyl Heptatrienyl, cyclooctyl, etc.; polycyclic carbocycles may include spiro, fused, and bridged carbocycles. Alicyclic groups can be substituted or unsubstituted.
  • the term "carbocyclyl" generally refers to a residue derived from a carbon atom having a carbocyclic ring by removing one hydrogen atom.
  • carbocycle generally refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon, the carbocycle contains 3 to 20 carbon atoms, may contain 3 to 12 carbon atoms, may contain 3 to 10 carbon atoms, may Contains 3 to 8 carbon atoms.
  • Non-limiting examples of monocyclic carbocycles include cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexene, cyclohexadiene, cycloheptane, cycloheptatriene, cyclooctane etc.; polycyclic carbocycles may include spiro, fused and bridged carbocycles. Carbocyclyl groups can be substituted or unsubstituted. Alicyclic and carbocyclic may be used interchangeably in some cases.
  • partially unsaturated generally refers to a cyclic structure containing at least one double or triple bond between the ring molecules.
  • the term “partially unsaturated” encompasses cyclic structures with multiple unsaturations, but is not intended to include aromatic or heteroaromatic rings as defined herein.
  • the term “unsaturated” means that the moiety has one or more degrees of unsaturation.
  • alicyclic group generally refers to a residue having two hydrogen atoms removed from a carbon atom of an alicyclic ring.
  • it can be cyclopropene, cyclobutane, cyclopentylene, cyclopentenylene, cyclohexylene, cyclohexenylene, cyclohexadienylene, cycloheptane cycloheptatrienyl, cyclooctylene, etc.
  • polycyclic carbocycles may include spiro, fused, and bridged carbocycles.
  • Alicyclic groups can be substituted or unsubstituted.
  • alicyclic heterocyclyl generally refers to stable non-aromatic 3- to 7-membered monocyclic carbocyclic structures, fused 7- to 10-membered bicyclic heterocyclic structures or bridged 6-membered -10-membered bicyclic heterocyclic structures, these cyclic structures can be either saturated or partially saturated, in addition to carbon atoms, these cyclic structures also contain one or more heteroatoms, wherein the heteroatoms can be selected from The following groups: oxygen, sulfur and nitrogen. For example, it contains 1-4 heteroatoms as defined above.
  • nitrogen when used to refer to an atom on an alicyclic ring structure may include nitrogen that has undergone a substitution reaction.
  • a heteroalicyclic group may include "heterocycloalkyl", which may refer to a stable non-aromatic 3- to 7-membered monocycloalkane structure, a fused 7- to 10-membered bicyclic heterocyclic structure or Bridged 6- to 10-membered bicyclic heterocyclic structures containing, in addition to carbon atoms, one or more heteroatoms, wherein the heteroatoms may be selected from the group consisting of oxygen, sulfur and nitrogen. For example, it contains 1-4 heteroatoms as defined above.
  • Heterocycloalkyl can be substituted or unsubstituted.
  • Alicyclic groups may be substituted or unsubstituted.
  • alicyclic heterocyclyl generally refers to a residue having two hydrogen atoms removed from a carbon atom of an alicyclic ring. Aliphatic heterocyclyl groups can be substituted or unsubstituted.
  • a heterocyclic group optionally substituted with an alkyl group means that an alkyl group may, but need not, be present, and the specification may include both instances where the heterocyclic group is substituted with an alkyl group and where the heterocyclic group is not substituted with an alkyl group. situation.
  • substituted generally means that one or more hydrogen atoms in a group, eg up to 5, eg 1 to 3 hydrogen atoms, independently of one another, are substituted by the corresponding number of substituents.
  • Substituents are only in their possible chemical positions, and those skilled in the art can determine (either experimentally or theoretically) possible or impossible substitutions without undue effort.
  • amino or hydroxyl groups with free hydrogens may be unstable when combined with carbon atoms with unsaturated (eg, olefinic) bonds.
  • alkyl alkenyl
  • cycloalkyl alkyl
  • C 3 alkyl C 3 -C 7 cycloalkoxy, C 1 -C 4 alkylcarbonylamino, etc.
  • C followed by a subscript number indicates the number of atoms present in the group number of carbon atoms.
  • C3 alkyl refers to an alkyl group having three carbon atoms (eg, n-propyl, isopropyl); in C1-10 , the members of the group can have any number falling within the range of 1-10 of carbon atoms.
  • One or more hydrogen atoms in the group are substituted by the corresponding number of substituents.
  • Substituents are only in their possible chemical positions, and those skilled in the art can determine (either experimentally or theoretically) possible or impossible substitutions without undue effort.
  • amino or hydroxyl groups with free hydrogens may be unstable when combined with carbon atoms with unsaturated (eg, olefinic) bonds.
  • the term "compound” generally refers to a substance having two or more different elements.
  • the compound of the present application may be an organic compound.
  • the compound of the present application may be a compound with a molecular weight of 500 or less, a compound with a molecular weight of 1,000 or less, or a compound with a molecular weight of 1,000 or more, or a compound of 10,000 or more and 100,000 or more. compound.
  • a compound may also refer to a compound connected by chemical bonds, for example, it may be a compound in which one or more molecules with a molecular weight below 1000 are connected with a biological macromolecule by chemical bonds, and the biological macromolecule may be a high polysaccharide, a protein , nucleic acids, peptides, etc.
  • the compounds of the present application may include compounds in which proteins are linked to one or more molecules with a molecular weight of less than 1000; A compound with less than 100,000 molecules linked together.
  • the term "about” generally refers to a range of 0.5%-10% above or below the specified value, such as 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10%.
  • the compounds of the present application include tautomers, mesomers, racemates, enantiomers, and/or diastereomers of the compounds.
  • the term “diastereomer” generally refers to stereoisomers that have two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers can have different physical properties, eg, melting points, boiling points, spectral properties, and reactivities.
  • the terms “tautomer” or “tautomeric form” are used interchangeably and generally refer to structural isomers of different energies that can be interconverted through a low energy barrier.
  • protontautomers also known as prototropic tautomers
  • prototropic tautomers include interconversions by migration of protons, such as keto-enol isomerization and imine-ene Amine isomerization.
  • Valence tautomers include interconversions by recombination of some of the bonding electrons.
  • mesome generally refers to atoms that contain asymmetry in the molecule, but have a symmetry factor such that the total optical rotation in the molecule is zero.
  • racemate or “racemic mixture” refers to a composition consisting of two enantiomeric species in equimolar amounts.
  • certain atoms of the compounds of the present application may occur in more than one isotopic form.
  • hydrogen may exist as protium ( 1 H), deuterium ( 2 H), and tritium ( 3 H), and carbon may exist naturally in three different isotopes ( 12 C, 13 C, and 14 C).
  • isotopes that can be incorporated into the compounds of the present application also include, but are not limited to, 15 N, 18 O, 17 O, 18 F, 32 P, 33 P, 129 I, 131 I, 123 I, 124 I, 125 I, or the like of isotopes.
  • the compounds of the present application may be enriched in one or more of these isotopes relative to their natural abundance.
  • Such isotopically enriched compounds can be used for a variety of purposes, as known to those skilled in the art.
  • substitution with heavy isotopes such as deuterium ( 2 H) may offer certain therapeutic advantages, which may be due to greater metabolic stability.
  • the natural abundance of deuterium ( 2 H) is about 0.015%. Therefore, for every 6,500 hydrogen atoms in nature, there is one deuterium atom. Accordingly, the deuterium-containing compounds of the present application have a deuterium abundance greater than 0.015% at one or more positions (as the case may be).
  • structures described herein may also include compounds that differ only in the presence or absence of one or more isotopically enriched atoms. For example, except that the hydrogen atom is replaced by deuterium or tritium, or the carbon atom is replaced by carbon 13 or carbon 14, the compounds whose structure is consistent with the present application are all within the scope of the present application.
  • the term "pharmaceutical composition” generally refers to a mixture containing one or more of the compounds described herein, or a physiologically/pharmaceutically acceptable salt or prodrug thereof, and other chemical components, as well as other Such as physiological/pharmaceutically acceptable carriers and excipients.
  • the pharmaceutical composition can facilitate the administration to the organism, facilitate the absorption of the active ingredient and then exert the biological activity.
  • the preparation of conventional pharmaceutical compositions can be found in the techniques commonly used in the art.
  • the term "pharmaceutically acceptable salt” or “pharmaceutically acceptable salt” generally refers to a salt of a compound or ligand-drug conjugate of the present application, or a salt of a compound described in the present application, Such salts can be safe and/or effective when used in mammals, and can have due biological activity, and the antibody-antibody drug conjugate compounds of the present application can form salts with acids, and the pharmaceutically acceptable salts are salts.
  • Non-limiting examples include: hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, citrate, acetate, succinate, ascorbate, oxalate, nitrate, pear Acid, hydrogen phosphate, dihydrogen phosphate, salicylate, hydrogen citrate, tartrate, maleate, fumarate, formate, benzoate, mesylate, ethyl acetate Sulfonate, benzenesulfonate, or p-toluenesulfonate.
  • conjugate generally refers to a compound of the present application prepared by one or more chemical reactions, or by one such as a bridge, spacer, or linking moiety, etc. or multiple connecting structures are connected to each other.
  • a pharmaceutically acceptable carrier generally refers to a carrier for administration of therapeutic agents, such as antibodies or polypeptides, genes and other therapeutic agents.
  • the term refers to any pharmaceutical carrier that itself does not induce the production of antibodies detrimental to the individual receiving the composition and can be administered without undue toxicity.
  • a pharmaceutically acceptable carrier can be distinguished from a nucleic acid vector used in genetic engineering to contain a gene of interest. Suitable carriers can be large, slowly metabolized macromolecules such as proteins, polysaccharides, polylactic acid, polyglycolic acid, polyamino acids, amino acid copolymers, lipid aggregates, and inactivated viral particles. Those skilled in the art are familiar with these vectors.
  • Pharmaceutically acceptable carriers in therapeutic compositions can include liquids such as water, saline, glycerol and ethanol. Auxiliary substances such as wetting or emulsifying agents, pH buffering substances and the like can also be present in these carriers.
  • Trop2 generally refer to a single-pass transmembrane type I cell membrane protein.
  • the term “Trop2” may also encompass homologues, variants and isoforms of Trop 2, including splice isoforms.
  • the term “Trop” also includes proteins having one or more sequences of Trop 2 homologues, variants and isoforms, as well as fragments of such sequences, so long as the variant protein (including isoforms).
  • Trop2 can be human Trop2.
  • Uniprot Accession No. P09758 provides a description of Trop2 and sequences.
  • HER2 generally refers to human epidermal growth factor receptor 2 (HER2).
  • HER2 refers to any native HER2 from any human source.
  • the term also encompasses "full-length” and unprocessed HER2 as well as any form of HER2 derived from processing in a cell (eg, the mature protein).
  • the term also encompasses naturally occurring variants and isoforms of HER2, such as splice variants or allelic variants.
  • Uniprot Accession No. P04626 provides a description of HER2 and sequences.
  • Nectin-4" generally refers to adhesion molecule 4.
  • Nectin-4" refers to any native Nectin-4 from any human source.
  • the term also encompasses "full-length” and unprocessed Nectin-4 as well as any form of Nectin-4 derived from processing in a cell (eg, mature protein).
  • the term also encompasses naturally occurring variants and isoforms of Nectin-4, such as splice variants or allelic variants.
  • Uniprot Accession No. Q96NY8 provides a description of Nectin-4 and sequences.
  • chimeric antibody generally refers to an antibody in which the variable region of a murine antibody is fused with the constant region of a human antibody, which can alleviate the immune response induced by the murine antibody.
  • a hybridoma that secretes a mouse-specific monoclonal antibody can be established, and then the variable region gene can be cloned from the mouse hybridoma cell, and the constant region gene of the human antibody can be cloned according to the needs.
  • the human constant region gene is connected into a chimeric gene and inserted into an expression vector, and the chimeric antibody molecule can be expressed in a eukaryotic system or a prokaryotic system.
  • humanized antibody also known as CDR-grafted antibody
  • CDR-grafted antibody generally refers to the grafting of murine CDR sequences into the framework of human antibody variable regions, i.e. different Types of human germline antibody framework sequences produced in antibodies.
  • the heterologous reaction induced by chimeric antibodies can be overcome because they carry a large amount of murine protein components.
  • framework sequences can be obtained from public DNA databases or published references that include germline antibody gene sequences.
  • the germline DNA sequences of the human heavy and light chain variable region genes can be found in the "VBase" human germline sequence database.
  • monoclonal antibodies has gone through four stages, namely: murine monoclonal antibodies, chimeric monoclonal antibodies, humanized monoclonal antibodies and fully human monoclonal antibodies.
  • the antibodies or ligands described herein may be fully human monoclonal antibodies.
  • Related technologies for the preparation of fully human antibodies include: human hybridoma technology, EBV transformation of B lymphocytes, phage display technology, transgenic mouse antibody preparation technology, and single B cell antibody preparation technology.
  • CDR generally refers to one of the six hypervariable regions within the variable domain of an antibody that primarily contribute to antigen binding.
  • One of the most commonly used definitions of the 6 CDRs is provided by Kabat E.A. et al., Chothia et al. and MacCallum et al.
  • the Kabat definition of CDRs can be applied to CDR1, CDR2 and CDR3 (CDR L1, CDR L2, CDR L3 or L1, L2, L3) of the light chain variable domains, as well as the heavy chain variable domains of CDR1, CDR2 and CDR3 (CDR H1, CDR H2, CDR H3 or H1, H2, H3).
  • group capable of coupling with a thiol group generally means that the compound A has a thiol group, the compound B has a group capable of coupling with a thiol group, and the compound B has a group capable of coupling with a thiol group through a group capable of coupling with a thiol group.
  • the group reacts with the sulfhydryl group of compound A, which can realize the connection between compound A and compound B.
  • linker usually refers to a chemical structural fragment or bond with one end connected to one group and the other end connected to another group. It can also be connected to other linkers and then to drugs and/or ligands. connected.
  • the direct or indirect linking of the ligand may refer to that the group is directly linked to the ligand through a covalent bond, or may be linked to the ligand through a linker.
  • the linker can be the structure shown by Q 1 described in this application.
  • chemical fragments comprising acid-labile linker structures (eg, hydrazones), protease-sensitive (eg, peptidase-sensitive) linker structures, photolabile linker structures, dimethyl linker structures, or disulfide-containing linker structures may be used or bond as a linker.
  • acid-labile linker structures eg, hydrazones
  • protease-sensitive linker structures eg, peptidase-sensitive linker structures
  • photolabile linker structures eg, dimethyl linker structures
  • disulfide-containing linker structures may be used or bond as a linker.
  • linking group generally refers to a group that has the ability to attach to another group.
  • the linking of the compound with another group can be achieved through a coupling reaction between the linking group and another group.
  • a maleimide group can serve as the linking group.
  • drug unit generally refers to a chemical moiety that directly or indirectly conjugates an antibody or antigen-binding fragment to form an immunoconjugate.
  • a drug unit includes, but is not limited to, compounds described herein with anti-tumor activity.
  • the drug unit includes a topoisomerase inhibitor.
  • the term "compound with antitumor activity” generally refers to a compound having the ability to reduce the proliferation rate, viability or metastatic activity of tumor cells.
  • anti-tumor activity may be manifested by a reduction in the growth rate of abnormal cells or by stabilization or shrinkage of tumor size during treatment, or by longer survival due to treatment compared to a control in the absence of treatment.
  • Antitumor activity can be assessed using recognized in vitro or in vivo tumor models, such as xenograft models.
  • the biologically active molecule in the conjugate is a compound with specific anti-tumor activity, for example, a radioisotope, such as At 211 , I 131 , I 125 , Y 90 , Re 186 , Re 188 , Sm 153 , Bi 212 , P 32 , Pb 212 , or radioisotopes of Lu; metal complexes, such as metal platinum complexes (such as oxaliplatin) or metal gold complexes; glycopeptide antibiotics, such as bleomycin or pingyangmycin; topoisomerase inhibitors; drugs that interfere with DNA synthesis, such as methotrexate, 5-fluorouracil, cytarabine, gemcitabine, mercaptopurine, pentostatin, fludarabine, cladrix Drugs acting on structural proteins, such as tubulin inhibitors (such as vinca alkaloids, vincristine, vinblastine, paclitaxel
  • topoisomerase inhibitor generally refers to compounds or derivatives thereof that include topoisomerase I inhibitors and topoisomerase II inhibitors.
  • topoisomerase I inhibitors include, but are not limited to, camptothecin and its analogs; topoisomerase II inhibitors (such as actinomycin D, doxorubicin, doxorubicin, docarmycin, daunorubicin, mitoxantrone, podophyllotoxin or etoposide, etc.).
  • a topoisomerase may refer to an enzyme that corrects the number of concatenated chains of DNA by cleaving phosphodiester bonds in one or both strands of DNA, followed by rewinding and sealing.
  • camptothecin analog generally refers to compounds that are structurally similar to or derived from camptothecin.
  • structure of camptothecin can be described in CAS Accession No. 7689-03-4.
  • a camptothecin analog may refer to ixatecan (Exatecan, CAS Accession No. 171335-80-1) or Belotecan (Belotecan, CAS Accession No. 256411-32-2).
  • non-camptothecin-type topoisomerase I inhibitors generally refers to indolecarbazoles, indenoisoquinolinones, triphenanthridines, and dibenzonaphthyridinones with topoisomerases I heterocyclic molecules with inhibitory activity mainly refer to Genz-644282 (CAS accession number 529488-28-6).
  • the term "expression-related" disease of a target generally means that the occurrence and/or progression of the disease is associated with the expression level of the target.
  • the expression level of a target in cells from a disease area such as within a particular tissue or organ of a patient, is increased, ie, highly expressed, relative to the expression level of normal cells from the tissue or organ.
  • the expression level of a certain target in cells from a disease area such as a particular tissue or organ of a patient, is reduced, ie, underexpressed, relative to the expression level of normal cells from the tissue or organ.
  • cells from a disease area such as within a particular tissue or organ of a patient, express a certain target, ie, are positive.
  • cells from a disease area such as a particular tissue or organ of a patient, do not express a certain target, ie, are negative.
  • target expression can be characterized by standard assays known in the art.
  • the term "effective amount” generally refers to the amount of a therapeutic agent that treats, ameliorates, or prevents a target disease or condition, or an amount that exhibits a measurable therapeutic or prophylactic effect.
  • the precise effective amount for a subject depends on the size and health of the subject, the nature and extent of the disorder, and the therapeutic agent and/or combination of therapeutic agents selected for administration. Therefore, it is useless to prespecify the exact effective amount. However, for a given situation, routine experimentation can be used to determine the effective amount, as is the judgment of the clinician.
  • each chiral carbon atom can optionally be in the R configuration or the S configuration, or a mixture of the R configuration and the S configuration.
  • the term "compound of the present application” refers to a compound of the present application.
  • the term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of the present application.
  • the tradename is intended to include the tradename product formulation, its corresponding generic drug, and the active pharmaceutical ingredient of the tradename product.
  • antibody is used in its broadest sense and specifically covers monoclonal antibodies, polyclonal antibodies, dimers, multimers, multispecific antibodies (eg, bispecific antibodies), and antibody fragments, so long as they are exhibit the desired biological activity.
  • Antibodies can be murine, human, humanized, chimeric, or derived from other species.
  • Antibodies are proteins produced by the immune system capable of recognizing and binding specific antigens.
  • Target antigens generally have a large number of binding sites, also referred to as epitopes, recognized by the CDRs of various antibodies. Each antibody that specifically binds to a different epitope has a different structure. Thus, an antigen can have more than one corresponding antibody.
  • Antibodies include full-length immunoglobulin molecules or immunologically active portions of full-length immunoglobulin molecules, i.e., molecules that contain an antigen or portion thereof that specifically binds a target of interest, including, but not limited to, cancer cells or Cells with autoimmune antibodies associated with autoimmune diseases.
  • the immunoglobulins described herein can be of any class (eg, IgG, IgE, IgM, IgD, and IgA), class (eg, IgGl, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass of immunoglobulin molecules. Immunoglobulins can be derived from any species.
  • the immunoglobulin is derived from human, murine or rabbit.
  • An "antibody fragment” may comprise a portion of a full-length antibody, typically its antigen-binding or variable region.
  • antibody fragments include: Fab, Fab', F(ab')2 and Fv fragments; diabodies; linear antibodies; minibodies; Fab expression library prepared fragments; anti-idiotypic (anti-Id) antibodies ; CDRs (complementarity determining regions); and any of the above epitope-binding fragments that immunospecifically bind to cancer cell antigens, viral antigens, or microbial antigens; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.
  • the antibody constituting the antibody-drug conjugate in the present application can maintain the antigen-binding ability of its original wild state.
  • the antibodies of the present application can, for example, specifically bind to an antigen.
  • Antigens involved include, for example, tumor-associated antigens (TAAs), cell surface receptor proteins and other cell surface molecules, regulators of cell survival, regulators of cell proliferation, molecules associated with tissue growth and differentiation (as known or predicted) functional), lymphokines, cytokines, molecules involved in the regulation of the cell cycle, molecules involved in angiogenesis, and molecules involved in angiogenesis (eg known antibodies bind antigens can be one or a subset of the above categories , while other subsets contain other molecules/antigens with specific properties (compared to the target antigen).
  • TAAs tumor-associated antigens
  • cell surface receptor proteins and other cell surface molecules regulators of cell survival, regulators of cell proliferation, molecules associated with tissue growth and differentiation (as known or predicted) functional
  • lymphokines cytokines
  • Antibodies used in antibody drug conjugates include, but are not limited to, targeting cell surface receptors and tumor-associated Antibodies to antigens.
  • tumor-associated antigens are well known in the industry and can be prepared by well-known antibody preparation methods and information in the industry. These targets can be specifically expressed on the surface of one or more cancer cells, while in a or a variety of non-cancerous cell surfaces expressed little or no.
  • tumor-associated polypeptides can be more overexpressed on the surface of cancer cells relative to the surface of non-cancer cells.
  • enrozumab generally refers to an antibody that targets Nectin-4.
  • Enfortumab can be described in WO2017042210A1.
  • ennozumab may refer to any antibody or antigen-binding fragment comprising the heavy chain variable region CDR1-3 and light chain variable region CDR1-3 of enrozumab.
  • ennozumab may refer to any antibody or antigen-binding fragment comprising the heavy chain variable region and light chain variable region of enrozumab.
  • Pertuzumab generally refers to an antibody that targets HER2.
  • Pertuzumab can be described in WO2014172371A2.
  • Pertuzumab may refer to any antibody or antigen-binding fragment comprising the heavy chain variable region CDR1-3 and the light chain variable region CDR1-3 of Pertuzumab.
  • Pertuzumab can refer to any antibody or antigen-binding fragment comprising the variable region of the heavy chain and the variable region of the light chain of Pertuzumab.
  • trastuzumab generally refers to an antibody that targets HER2.
  • Trastuzumab can be described in US20060275305A1.
  • trastuzumab may refer to any antibody or antigen-binding fragment comprising the heavy chain variable region CDR1-3 and light chain variable region CDR1-3 of trastuzumab.
  • trastuzumab may refer to any antibody or antigen-binding fragment comprising the heavy and light chain variable regions of trastuzumab.
  • certolizumab generally refers to an antibody that targets TROP2.
  • Sacituzumab hRS7
  • certolizumab may refer to any antibody or antigen-binding fragment comprising the heavy chain variable region CDR1-3 and light chain variable region CDR1-3 of certolizumab.
  • certolizumab may refer to any antibody or antigen-binding fragment comprising the heavy and light chain variable regions of certolizumab.
  • Patritumab generally refers to an antibody that targets HER3.
  • Patritumab can be described in CN102174105B.
  • Patritumab monoclonal antibody may refer to any antibody or antigen-binding fragment comprising the heavy chain variable region CDR1-3 and light chain variable region CDR1-3 of Patritumab.
  • Patritumab may refer to any antibody or antigen-binding fragment comprising the variable region of the heavy chain and the variable region of the light chain of Patritumab.
  • the term "antibody H01L02" generally refers to an antibody targeting CDH6.
  • the monoclonal antibody H01L02 can be described in WO2018212136, US20200171163A1.
  • the H01L02 mAb can be the mAb used by the drug DS6000.
  • the H01L02 monoclonal antibody may refer to any antibody or antigen-binding fragment comprising the heavy chain variable region CDR1-3 and the light chain variable region CDR1-3 of the H01L02 monoclonal antibody.
  • the H01L02 mAb can refer to any antibody or antigen-binding fragment comprising the heavy chain variable region and the light chain variable region of the H01L02 mAb.
  • polypeptide residue generally refers to a residue comprising one or more amino acid residues linked together.
  • one or more amino acids in a polypeptide residue may be optionally substituted.
  • the polypeptide residues of the present application may be selected from the group consisting of phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala), valine-citrulline (Val) -Cit), glutamic acid-valine-alanine (Glu-Val-Ala), glutamic acid-valine-citrulline (Glu-Val-Cit), valine-lysine ( Val-Lys), Alanine-Alanine-Alanine (Ala-Ala-Ala), Alanine-Alanine-Asparagine (Ala-Ala-Asn), and Glycine-Glycine-Phenylalanine Acid-glycine (Gly-Gly-Phe-Gly).
  • polyethylene glycol generally refers to a residue comprising one or more ethylene glycol residues linked together.
  • a polyethylene glycol group may comprise - ( CH2CH2O ) p- , where p is a number of at least one.
  • polyethylene glycol groups in this application may be optionally substituted.
  • glycol group generally refers to a polyethylene glycol group.
  • glycol groups in this application may be optionally substituted.
  • a number preceding a glycol group may indicate the number of ethylene glycol units of a glycol group, eg, a diethylene glycol group may refer to the polymerized residue of two glycols.
  • polysarcosine residue generally refers to a residue comprising one or more sarcosine residues linked together.
  • a polysarcosine residue may comprise -(COCH2N( CH3 ) ) q- , where q is a number of at least one.
  • polysarcosine residues in the present application may be optionally substituted.
  • a structure containing polysarcosine residues can be where n2 is a number from 4 to 18.
  • sodium dodecyl sulfate polyacrylamide gel electrophoresis generally refers to an analytical characterization technique for substances.
  • sodium dodecyl sulfate polyacrylamide gel electrophoresis SDS-PAGE
  • SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis
  • hydrophobic chromatography generally refers to an analytical technique based on differences in the hydrophobicity of substances.
  • liquid phase mass spectrometry generally refers to an analytical method for identifying constituents of matter.
  • liquid chromatography-mass spectrometry can analyze the molecular weight of the substance to be tested by liquid chromatography-mass spectrometry.
  • tumor generally refers to any new pathological tissue proliferation.
  • angiogenesis is part of the tumor profile.
  • Tumors can be benign or malignant.
  • the term “tumor” is generally used to refer to benign or malignant tumors, while the term “cancer” is generally used to refer to malignant tumors, which may be metastatic or non-metastatic.
  • Tumors diagnosable by the methods of the present application are selected from the group consisting of breast cancer, ovarian cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, acute lymphoblastic leukemia, anaplastic large cell lymphoma, multiple myeloma, Prostate cancer, non-small cell lung cancer, small cell lung cancer, malignant melanoma, squamous cell carcinoma, glioblastoma, renal cell carcinoma, gastrointestinal tumors, pancreatic cancer, prostate cancer, rectal colon, gastric cancer, glial tumor and mesothelioma.
  • these tissues can be isolated from readily available sources by methods well known to those skilled in the art.
  • the application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable compound thereof.
  • An acceptable salt, prodrug or solvate, wherein the compound may comprise the structure of formula (C-Trop-2):
  • Q 1 may contain a linker
  • L 1a can be selected from the following group: optionally substituted alkylene, optionally substituted polyethylene glycol, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted idene cyclyl, optionally substituted alicyclic heterocyclyl, optionally substituted arylene, and optionally substituted heteroarylene;
  • L2 may comprise optionally substituted polypeptide residues
  • L 3 may contain an optionally substituted spacer group, for example, the spacer group of the present application may be self-degradable.
  • the spacer groups of the present application may comprise optionally substituted or optionally substituted
  • L2 and/or L3 may comprise optionally substituted polysarcosine residues
  • T may contain drug units
  • Ab can be a ligand capable of binding Trop-2, and m can be a number from 1-8.
  • the present application also provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or a mixture thereof A pharmaceutically acceptable salt, prodrug or solvate, wherein the compound may comprise a structure represented by the formula (C-Trop-2-M):
  • Q 1 may contain a linker
  • L 1a can be selected from the following group: optionally substituted alkylene, optionally substituted polyethylene glycol, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted idene cyclyl, optionally substituted alicyclic heterocyclyl, optionally substituted arylene, and optionally substituted heteroarylene;
  • L2 may comprise optionally substituted polypeptide residues
  • L 3 may contain an optionally substituted spacer group, for example, the spacer group of the present application may be self-degradable.
  • the spacer groups of the present application may comprise optionally substituted or optionally substituted
  • L 2 and/or L 3 may comprise optionally substituted structural unit-X
  • T may contain drug units
  • Ab can be a ligand capable of binding Trop-2, and m can be a number from 1-8.
  • L is selected from the group of: optionally substituted and optionally substituted
  • the benzene ring of L 3 may be substituted by the optionally substituted structural unit -X.
  • the structural unit -X can be selected from the group of: optionally substituted wherein X 1 is selected from the group consisting of carbonyl, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight chain heteroalkyl containing 1-8 atoms, and Linear-cyclic heteroalkyl groups containing 1-8 atoms containing 1-3 atoms selected from N, O or S ; wherein X is selected from the group consisting of hydrogen, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkane containing 1-8 atoms group, the heteroalkyl group contains 1-3 atoms selected from N, O or S; wherein X 3 is
  • the benzene ring of L 3 may be substituted by the optionally substituted structural unit -X.
  • the building block -X may contain an optionally substituted wherein X 1 is selected from the group consisting of C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight chain heteroalkyl containing 1-8 atoms, and containing 1 -8-atom straight-chain-cyclic heteroalkyl, said heteroalkyl containing 1-3 atoms selected from N, O or S, said C 1 -C 8 alkyl, C 1 -C 8 Alkoxy, C1 - C6cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkyl containing 1-8 atoms are each independently optionally selected from Substituted with one or more substituents of deuterated, halogen, cyano, nitro, amino, alkyl, carboxyl, al
  • the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C-Trop-2-M2):
  • L can contain an optionally substituted alkylene group, an optionally substituted polyethylene glycol group and an optionally substituted alicyclic group;
  • R 1 can be an optionally substituted isopropyl group or an optionally substituted benzyl group ;
  • R 2 can be optionally substituted methyl, optionally substituted or optionally substituted
  • R 3 can be hydrogen or R 5 ; wherein R 5 can be optionally substituted T may comprise ixatecan (Exatecan, CAS accession number 171335-80-1) and/or belonotecan (Belotecan, CAS accession number 256411-32-2) and/or Genz-644282 (CAS accession number 529488- 28-6).
  • Q 1 may comprise a linker coupled with a thiol group.
  • Q 1 can be selected from the group of: optionally substituted optionally substituted optionally substituted and optionally substituted
  • L 1a may be selected from the group consisting of optionally substituted C 1 -C 7 alkylene, optionally substituted diethylene glycol to octaethylene glycol, optionally substituted C 3 -C 6 alkylene Alicyclic, optionally substituted arylene, and optionally substituted heteroarylene.
  • L 1a may be selected from the group consisting of optionally substituted methylene, optionally substituted ethylene, optionally substituted propylene, optionally substituted butylene, optionally substituted pentylene , optionally substituted diethylene glycol, optionally substituted tetraethylene glycol, optionally substituted hexaethylene glycol, optionally substituted octaethylene glycol, and optionally substituted cyclohexylene.
  • L 1a can be selected from the group consisting of optionally substituted methylene, optionally substituted ethylene, optionally substituted propylene, optionally substituted butylene, and optionally substituted pentylene base.
  • L 1a can be selected from the group of: optionally substituted diethylene glycol, optionally substituted triethylene glycol, optionally substituted tetraethylene glycol, optionally substituted pentaethylene glycol, optionally substituted Substituted hexaethylene glycol, optionally substituted heptaethylene glycol, and optionally substituted octaethylene glycol.
  • L 1a may be selected from the group consisting of optionally substituted cyclopropylene, optionally substituted cyclobutylene, and optionally substituted cyclohexylene.
  • L2 may comprise optionally substituted polypeptide residues consisting of amino acids selected from the group consisting of phenylalanine, isoleucine, leucine, tryptophan, valine, methionine , tyrosine, alanine, threonine, histidine, serine, glutamine, arginine, lysine, asparagine, glutamic acid, proline, citrulline, aspartate acid and glycine.
  • amino acids selected from the group consisting of phenylalanine, isoleucine, leucine, tryptophan, valine, methionine , tyrosine, alanine, threonine, histidine, serine, glutamine, arginine, lysine, asparagine, glutamic acid, proline, citrulline, aspartate acid and glycine.
  • L 2 may comprise optionally substituted polypeptide residues consisting of amino acids selected from the group consisting of glycine, phenylalanine, valine, alanine, arginine, citrulline, aspartate acid, asparagine and lysine.
  • L 2 may comprise an optionally substituted polypeptide residue selected from the group consisting of: phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala), valine Acid-citrulline (Val-Cit), glutamic acid-valine-alanine (Glu-Val-Ala), glutamic acid-valine-citrulline (Glu-Val-Cit), valine Amino-Lysine (Val-Lys), Alanine-Alanine-Alanine (Ala-Ala-Ala), Alanine-Alanine-Asparagine (Ala-Ala-Asn) and Glycine-Glycine-Phenylalanine-Glycine (Gly-Gly-Phe-Gly).
  • Phe-Lys phenylalanine-lysine
  • Val-Ala valine-alanine
  • Val-Cit valine Acid-citrulline
  • Glu-Val-Ala glutamic acid-
  • L 2 may comprise an optionally substituted polypeptide residue selected from the group consisting of: phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala), valine Acid-citrulline (Val-Cit) and valine-lysine (Val-Lys).
  • L2 comprises a lysine residue
  • the lysine residue can be substituted by a structure R1 comprising a polysarcosine residue.
  • any H contained in L 2 may be substituted by R 1 .
  • the R 1 can be optionally substituted wherein n1 is a number from 4 to 18, and R is selected from the group consisting of C 1 -C 6 alkyl, C 1 -C 6 cycloalkyl, and C 1 -C 6 alkoxy.
  • n1 can be 4 to 18, 8 to 18, 4 to 12, or 8 to 12.
  • n1 can be 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18.
  • L is selected from the group of: optionally substituted and optionally substituted
  • the benzene ring of L3 can be substituted with a structure R2 comprising a polysarcosine residue.
  • any H contained in the benzene ring of L 3 may be substituted by R 2 .
  • the structural unit -X- is selected from the group consisting of: optionally substituted wherein X 1 is selected from the group consisting of carbonyl, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight chain heteroalkyl containing 1-8 atoms, and Linear-cyclic heteroalkyl groups containing 1-8 atoms containing 1-3 atoms selected from N, O or S ; wherein X is selected from the group consisting of hydrogen, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkane containing 1-8 atoms group, the heteroalkyl group contains 1-3 atoms selected from N, O or S;
  • the benzene ring of L 3 is connected to the optionally substituted polysarcosine residue through a structural unit -X-, which is selected from, but is not limited to: wherein X 1 is selected from the group consisting of C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight chain heteroalkyl containing 1-8 atoms, and containing 1 -8-atom straight-chain-cyclic heteroalkyl, said heteroalkyl containing 1-3 atoms selected from N, O or S, said C 1 -C 8 alkyl, C 1 -C 8 Alkoxy, C1 - C6cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkyl containing 1-8 atoms are each independently optionally selected from Substituted with one or more substituents of deuterated, halogen, cyano, nitro, amino, alkyl, carboxyl,
  • the optionally substituted polysarcosine residue comprises wherein n2 is a number from 4 to 18, and R is selected from the group consisting of C 1 -C 6 alkyl, C 1 -C 6 cycloalkyl, and C 1 -C 6 alkoxy.
  • the structural unit -X- is selected from the group consisting of optionally substituted wherein X 1 is selected from the group consisting of carbonyl, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight chain heteroalkyl containing 1-8 atoms, and Linear-cyclic heteroalkyl groups containing 1-8 atoms containing 1-3 atoms selected from N, O or S ; wherein X is selected from the group consisting of hydrogen, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkane containing 1-8 atoms group, the heteroalkyl group contains 1-3 atoms selected from N, O or S; wherein X 3 is a covalent bond or is selected from the following group: hydrogen, C 1 -C 8 alkyl, C 1 -
  • the optionally substituted polysarcosine residue comprises wherein n2 is a number from 4 to 18, and R is selected from the group consisting of C 1 -C 6 alkyl, C 1 -C 6 cycloalkyl, and C 1 -C 6 alkoxy.
  • the structural unit -X- is selected from the group consisting of optionally substituted wherein X 1 is selected from the group consisting of carbonyl, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight chain heteroalkyl containing 1-8 atoms, and Linear-cyclic heteroalkyl groups containing 1-8 atoms containing 1-3 atoms selected from N, O or S ; wherein X is selected from the group consisting of hydrogen, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkane containing 1-8 atoms group, the heteroalkyl group contains 1-3 atoms selected from N, O or S; wherein X 3 is a covalent bond or is selected from the following group: hydrogen, C 1 -C 8 alkyl, C 1 -
  • n2 can be 4 to 18, 8 to 18, 4 to 12, or 8 to 12.
  • n2 can be 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18.
  • T may comprise a compound having antitumor activity.
  • T can comprise a topoisomerase inhibitor
  • T can include camptothecin-based and non-camptothecin-based topoisomerase I inhibitors.
  • T may comprise ixatecan (Exatecan, CAS accession number 171335-80-1) and/or belotecan (Belotecan, CAS accession number 256411-32-2) and/or Genz-644282 (CAS accession number 256411-32-2) Accession No. 529488-28-6).
  • T can be selected from the following group:
  • the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C-Trop-2):
  • Q 1 may comprise a linker coupled with a thiol group
  • L2 may comprise optionally substituted polypeptide residues consisting of amino acids selected from the group consisting of glycine, phenylalanine, valine, alanine, arginine, citrulline, aspartic acid, aspartic acid amide and lysine,
  • L 3 may contain optionally substituted
  • L 2 may comprise a polysarcosine residue
  • T may contain a topoisomerase inhibitor.
  • the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C-Trop-2):
  • Q 1 may comprise a linker coupled with a thiol group
  • L2 may comprise optionally substituted polypeptide residues consisting of amino acids selected from the group consisting of glycine, phenylalanine, valine, alanine, arginine, citrulline, aspartic acid, aspartic acid amide and lysine,
  • L 3 may contain optionally substituted
  • L3 may comprise a polysarcosine residue
  • T may contain a topoisomerase inhibitor.
  • the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C-Trop-2):
  • Q 1 may contain optionally substituted
  • L 2 may comprise a structure selected from the group consisting of phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala), valine-citrulline (Val-Cit) and valine-lysine (Val-Lys),
  • L 3 may contain optionally substituted
  • L2 may comprise a lysine residue, and the lysine residue may be optionally substituted Substitution, where n1 is a number from 4 to 18,
  • T may comprise ixatecan (Exatecan, CAS accession number 171335-80-1) and/or belonotecan (Belotecan, CAS accession number 256411-32-2) and/or Genz-644282 (CAS accession number 529488- 28-6).
  • the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C-Trop-2):
  • Q 1 can be selected from the following group: optionally substituted optionally substituted optionally substituted and optionally substituted
  • L 2 may comprise a structure selected from the group consisting of phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala), valine-citrulline (Val-Cit) and valine-lysine (Val-Lys),
  • L 3 may contain optionally substituted
  • L can be optionally substituted Substitution, wherein L can be optionally substituted Substitution, wherein n2 is a number from 4 to 18,
  • T may comprise ixatecan (Exatecan, CAS accession number 171335-80-1) and/or belonotecan (Belotecan, CAS accession number 256411-32-2) and/or Genz-644282 (CAS accession number 529488- 28-6).
  • the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C2-Trop-2):
  • L can contain an optionally substituted alkylene group, an optionally substituted polyethylene glycol group and an optionally substituted alicyclic group;
  • R 1 can be an optionally substituted isopropyl group or an optionally substituted benzyl group ;
  • R 2 can be optionally substituted methyl, optionally substituted or optionally substituted R 3 can be hydrogen or optionally substituted methyl;
  • R 2 can be optionally substituted Substitute, where n1 is a number from 4 to 18, and T may comprise ixatecan (Exatecan, CAS Accession No. 171335-80-1) and/or Belotecan (Belotecan, CAS Accession No. 256411-32-2) and /or Genz-644282 (CAS Accession No. 529488-28-6).
  • the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C2-Trop-2):
  • L can contain an optionally substituted alkylene group, an optionally substituted polyethylene glycol group and an optionally substituted alicyclic group;
  • R 1 can be an optionally substituted isopropyl group or an optionally substituted benzyl group ;
  • R 2 can be optionally substituted methyl, optionally substituted or optionally substituted R 3 can be hydrogen or optionally substituted methyl;
  • R 2 can be optionally substituted Substitute, where n2 is a number from 4 to 18, and T may comprise ixatecan (Exatecan, CAS Accession No. 171335-80-1) and/or Belotecan (Belotecan, CAS Accession No. 256411-32-2) and /or Genz-644282 (CAS Accession No. 529488-28-6).
  • the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C2-Trop-2):
  • L can contain an optionally substituted alkylene group, an optionally substituted polyethylene glycol group and an optionally substituted alicyclic group;
  • R 1 can be an optionally substituted isopropyl group or an optionally substituted benzyl group ;
  • R 2 can be optionally substituted R 3 can be hydrogen or optionally substituted methyl;
  • R 4 can be optionally substituted where n1 is a number from 4 to 18, and T may contain ixatecan (Exatecan, CAS accession number 171335-80-1) and/or belotecan (Belotecan, CAS accession number 256411-32-2) and/or Genz-644282 (CAS Accession No. 529488-28-6).
  • the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C2-Trop-2):
  • L can contain an optionally substituted alkylene group, an optionally substituted polyethylene glycol group and an optionally substituted alicyclic group;
  • R 1 can be an optionally substituted isopropyl group or an optionally substituted benzyl group ;
  • R 2 can be optionally substituted methyl, optionally substituted or optionally substituted
  • R 3 can be hydrogen or R 5 ; wherein R 5 can be optionally substituted where n2 is a number from 4 to 18, and T may comprise ixatecan (Exatecan, CAS accession number 171335-80-1) and/or belotecan (Belotecan, CAS accession number 256411-32-2) and/or Genz-644282 (CAS Accession No. 529488-28-6).
  • the Ab can comprise an anti-Trop-2 antibody or antigen-binding fragment thereof.
  • the antibody may be selected from the group consisting of murine antibodies, chimeric antibodies, humanized antibodies, and fully human antibodies.
  • the antibody may comprise a monoclonal antibody.
  • the antibody may comprise a bispecific antibody.
  • the antigen-binding fragment may be selected from the group consisting of Fab, Fab', Fv fragments, F(ab') 2 , F(ab) 2 , scFv, di-scFv, VHH and dAb.
  • the heavy chains HCDR1, HCDR2 and HCDR3 and light chains LCDR1, LCDR2 and LCDR3 of the Ab comprise heavy chains HCDR1, HCDR2 and HCDR3 and light chains LCDR1, LCDR2 and LCDR3, respectively, of an anti-Trop-2 antibody.
  • the heavy chain variable region VH and light chain variable region VL of the Ab comprise the heavy chain variable region VH and light chain variable region VL, respectively, of an anti-Trop-2 antibody.
  • the heavy and light chains of the Ab comprise the heavy and light chains, respectively, of an anti-Trop-2 antibody.
  • the Ab can comprise certolizumab (Sacituzumab, hRS7).
  • m can be determined by a method selected from the group consisting of hydrophobic chromatography, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and liquid phase mass spectrometry.
  • m is the average value of the molar ratio of drug molecules to monoclonal antibody molecules in the antibody-conjugated drug obtained by coupling a single monoclonal antibody molecule with a cytotoxic drug, and m can be an integer or decimal from 1 to 8, such as , m can be about 1 to about 2, about 1 to about 3, about 1 to about 4, about 1 to about 5, about 1 to about 6, about 1 to about 7, or about 1 to about 8; for example, m can be about 2 to about 8, about 3 to about 8, about 4 to about 8, about 5 to about 8, about 6 to about 8, about 7 to about 8, or about 1, about 2, about 3, about 4, About 5, about 6, about 7 or about 8.
  • the present application also provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or a mixture thereof
  • a pharmaceutically acceptable salt, prodrug or solvate
  • the compound comprises a structure selected from the group consisting of:
  • Ab is a ligand capable of binding Trop-2, and m is a number from 1 to 8.
  • the ligands described herein may be protein hormones, lectins, growth factors, antibodies, or other molecules capable of binding to cells, receptors and/or antigens.
  • the ligand of the present application can be an anti-Trop-2 antibody or an antigen-binding fragment thereof.
  • the ligand comprises at least one CDR in the variable region VL of the antibody light chain.
  • the CDRs described in the present application may be defined according to Kabat; or may be defined according to Chothia, and the CDR sequences defined in various ways are all included within the protection scope of the present application.
  • the antigen-binding protein of the present application may comprise CDR1-3 of the heavy chain variable region and CDR1-3 of the light chain variable region, wherein CDR1-3 of the heavy chain variable region and CDR1-3 of the light chain variable region 3 can be CDR1-3 of the heavy chain variable region and CDR1-3 of the light chain variable region of Sacituzumab, respectively.
  • the antigen-binding protein of the present application may have the ability to bind to TROP2.
  • the antigen binding protein of the present application can be a heavy chain variable region comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region and the light chain variable region can be the heavy chain variable region of Sacituzumab, respectively variable region and light chain variable region.
  • the antigen-binding protein of the present application may have the ability to bind to TROP2.
  • the antigen binding proteins of the present application may be comprised of heavy and light chains, wherein the heavy and light chains may be the heavy and light chains of Sacituzumab, respectively.
  • the heavy chain amino acid sequence of Sacituzumab can be shown as SEQ ID NO:1
  • the light chain amino acid sequence of Sacituzumab can be shown as SEQ ID NO:2.
  • the ligands of the present application can be: for example, the heavy chain amino acid sequence of Sacituzumab can be as shown in SEQ ID NO: 1, ) light chain amino acid sequence can be as shown in SEQ ID NO:2.
  • the heavy chain amino acid sequence of Trastuzumab can be shown in SEQ ID NO:3, and the light chain amino acid sequence of Trastuzumab can be shown in SEQ ID NO:4.
  • the heavy chain amino acid sequence of Pertuzumab can be shown in SEQ ID NO:5, and the light chain amino acid sequence of Pertuzumab can be shown in SEQ ID NO:6.
  • the heavy chain amino acid sequence of Enfortumab can be shown in SEQ ID NO:7, and the light chain amino acid sequence of Enfortumab can be shown in SEQ ID NO:8.
  • the heavy chain amino acid sequence of Patritumab can be set forth in SEQ ID NO:9, and the light chain amino acid sequence of Patritumab can be set forth in SEQ ID NO:10.
  • the heavy chain amino acid sequence of antibody H01L02 can be set forth in SEQ ID NO:11, and the light chain amino acid sequence of antibody H01L02 can be set forth in SEQ ID NO:12.
  • Antibodies of the present application can be prepared using techniques well known in the art, such as hybridoma methods, recombinant DNA techniques, phage display techniques, synthetic techniques, or a combination of these techniques, or other techniques known in the art.
  • Variants may refer to amino acid sequence mutants of antibodies, as well as covalent derivatives of the native polypeptide, provided that biological activity comparable to the native polypeptide is retained.
  • Amino acid sequence mutants generally differ from the native amino acid sequence by the substitution of one or more amino acids in the native amino acid sequence or the deletion and/or insertion of one or more amino acids in the polypeptide sequence.
  • Deletion mutants include fragments of the native polypeptide and N-terminal and/or C-terminal truncation mutants. Typically the amino acid sequence mutants have at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% or more homology compared to the native sequence.
  • the drugs can be released into cells in an active form through the endocytosis of the conjugates or drug infiltration.
  • the antibody-drug conjugate of the present application can be used to treat a target disease, and the antibody-drug conjugate of the present application can be administered to a subject (eg, a human) in a therapeutically effective amount through a suitable route.
  • a subject in need of treatment may be a patient at risk, or suspected of having a disorder associated with the activity or level of expression of a particular antigen. Such patients can be identified by routine physical examination.
  • delivery can be performed by methods routine in the art. For example, it can be introduced into cells by using liposomes, hydrogels, cyclodextrins, biodegradable nanocapsules, or bioadhesive microspheres.
  • the nucleic acid or vector can be delivered locally by direct injection or by using an infusion pump.
  • Other methods may include the use of various transport and carrier systems through the use of conjugates and biodegradable polymers.
  • the application provides a pharmaceutical composition, which may contain the compound described in any one of the application, or a tautomer, meso, racemate, enantiomer, The diastereomers, or mixtures thereof, or pharmaceutically acceptable salts, prodrugs or solvates thereof, and may optionally contain a pharmaceutically acceptable carrier.
  • the pharmaceutical composition described in the application may contain one or more excipients, which may be selected from the following group of ingredients: fillers (diluents), binders, wetting agents, disintegrating agents and excipients, etc.
  • the composition may contain from 0.1 to 99% by weight of active compound.
  • compositions containing the active ingredient may be in a form suitable for oral administration, such as tablets, dragees, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups.
  • Oral compositions may be prepared according to any method known in the art for the preparation of pharmaceutical compositions, which may contain binders, fillers, lubricants, disintegrants or pharmaceutically acceptable wetting agents, etc.
  • the composition may also contain one or more ingredients selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preservatives.
  • Aqueous suspensions may contain the active materials in admixture with excipients suitable for the preparation of aqueous suspensions.
  • the aqueous suspensions may also contain one or more preservatives, such as one or more coloring agents, one or more flavoring agents, and one or more sweetening agents.
  • Oily suspensions can be formulated by suspending the active ingredient in vegetable oils.
  • the oily suspensions may contain thickening agents. The above-mentioned sweetening and flavoring agents may also be added.
  • the pharmaceutical compositions may also provide the active ingredient as dispersible powders and granules for preparation of aqueous suspensions by admixing with water one or more of a dispersing agent, wetting agent, suspending agent or preservative. Other excipients such as sweetening, flavouring and colouring agents may also be added. These compositions are preserved by the addition of antioxidants such as ascorbic acid.
  • the pharmaceutical compositions of the present application may also be in the form of oil-in-water emulsions.
  • the pharmaceutical compositions may be in the form of sterile injectable aqueous solutions.
  • acceptable vehicles or solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution.
  • the sterile injectable preparation may be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in an oily phase.
  • the active ingredient is dissolved in a mixture of soybean oil and lecithin.
  • the oil solution can then be processed to form a microemulsion by adding it to a mixture of water and glycerol. Injections or microemulsions can be injected into a patient's bloodstream by local bolus injection.
  • solutions and microemulsions can be administered in a manner that maintains a constant circulating concentration of the compounds of the application.
  • a continuous intravenous drug delivery device can be used.
  • the device may be an intravenous pump.
  • compositions may be in the form of sterile injectable aqueous or oily suspensions for intramuscular and subcutaneous administration.
  • This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned herein above.
  • the sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent. Alternatively, sterile fixed oils are conveniently employed as a solvent or suspending medium.
  • the compounds of the present application may be administered in the form of suppositories for rectal administration.
  • These pharmaceutical compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid in the rectum and therefore will melt in the rectum to release the drug.
  • suitable non-irritating excipient include cocoa butter, glycerinated gelatin, hydrogenated vegetable oils, polyethylene glycols of various molecular weights and mixtures of fatty acid esters of polyethylene glycols.
  • the dosage of a drug to be administered depends on a variety of factors including, but not limited to, the following factors: the activity of the particular compound used, the age of the patient, the weight of the patient, the health of the patient, the behavior of the patient , the patient's diet, administration time, administration mode, excretion rate, combination of drugs, etc.; in addition, the optimal treatment mode such as the mode of treatment, the compound or its tautomer, meso isomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, and/or compounds or tautomers, mesoisomers,
  • the daily amount of racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, or the species of pharmaceutically acceptable salts can be verified according to conventional therapeutic regimens.
  • the pharmaceutical composition of the present application may contain a safe and effective amount of the antibody-drug conjugate of the present application and a pharmaceutically acceptable carrier.
  • a pharmaceutically acceptable carrier can include, but are not limited to, saline, buffers, dextrose, water, glycerol, ethanol, and combinations thereof.
  • the pharmaceutical preparation should match the mode of administration, and the pharmaceutical composition of the present application can be prepared in the form of a solution, for example, prepared by a conventional method with a physiological saline or an aqueous solution containing glucose and other adjuvants.
  • the pharmaceutical composition can be manufactured under sterile conditions.
  • the active ingredient may be administered in a therapeutically effective amount.
  • the effective amount of the antibody-drug conjugates described herein may vary with the mode of administration, the severity of the disease to be treated, and the like. Selection of an effective amount can be determined by one of ordinary skill in the art based on various factors (eg, through clinical trials). The factors may include, but are not limited to: the pharmacokinetic parameters of the diabody conjugate such as bioavailability, metabolism, half-life, etc.; the severity of the disease to be treated by the patient, the weight of the patient, the immune status, route of administration, etc. Generally, when the antibody-drug conjugates of the present application are administered in an appropriate dose per day, satisfactory results can be obtained. For example, several divided doses may be administered daily or the dose may be proportionally reduced as dictated by the exigencies of the therapeutic situation.
  • the compounds of the present application may be administered alone, or may be administered in combination with other pharmaceutically acceptable therapeutic agents.
  • a safe and effective amount of the compound of the present application can be applied to a mammal (such as a human) in need of treatment, and the dose can be a pharmaceutically effective dose when administered, and the specific dose can also be considered. factors such as route, patient's health status, etc.
  • the present application provides a compound of the present application, or a tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable form thereof
  • the tumor can be selected from tumors associated with expression of the following group of targets: HER2.
  • the tumor associated with the expression of the target includes a tumor with high expression of the target and/or a tumor positive for the target.
  • the tumor comprises a solid tumor and/or a hematological tumor.
  • the tumor is selected from the group consisting of breast cancer, ovarian cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, acute lymphoblastic leukemia, anaplastic large cell lymphoma, multiple myeloma, prostate cancer, Non-small cell lung cancer, small cell lung cancer, malignant melanoma, squamous cell carcinoma, glioblastoma, renal cell carcinoma, gastrointestinal tumors, pancreatic cancer, prostate cancer, rectal cancer, gastric cancer, glioma and skin tumor.
  • the present application provides a compound of the present application, or a tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable form thereof Acceptable salts, prodrugs or solvates, and/or pharmaceutical compositions of the present application, which can be used for use in medicaments for the treatment and/or prevention of tumors.
  • the tumor can be selected from tumors associated with expression of the following group of targets: HER2.
  • the tumor associated with the expression of the target includes a tumor with high expression of the target and/or a tumor positive for the target.
  • the tumor comprises a solid tumor and/or a hematological tumor.
  • the tumor is selected from the group consisting of breast cancer, ovarian cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, acute lymphoblastic leukemia, anaplastic large cell lymphoma, multiple myeloma, prostate cancer, Non-small cell lung cancer, small cell lung cancer, malignant melanoma, squamous cell carcinoma, glioblastoma, renal cell carcinoma, gastrointestinal tumors, pancreatic cancer, prostate cancer, rectal cancer, gastric cancer, glioma and skin tumor.
  • the present application provides a method of preventing and/or treating tumors, which may include administering to a subject a compound of the present application, or a tautomer, meso, racemate, enantiomer, A diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, and/or the pharmaceutical composition of the present application.
  • the tumor can be selected from tumors associated with expression of the following group of targets: HER2.
  • the tumor associated with the expression of the target includes a tumor with high expression of the target and/or a tumor positive for the target.
  • the tumor comprises a solid tumor and/or a hematological tumor.
  • the tumor is selected from the group consisting of breast cancer, ovarian cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, acute lymphoblastic leukemia, anaplastic large cell lymphoma, multiple myeloma, prostate cancer, Non-small cell lung cancer, small cell lung cancer, malignant melanoma, squamous cell carcinoma, glioblastoma, renal cell carcinoma, gastrointestinal tumors, pancreatic cancer, prostate cancer, rectal cancer, gastric cancer, glioma and skin tumor.
  • Fmoc is 9-fluorenylmethoxycarbonyl protecting group (9-fluorenylmethyloxycarbonyl)
  • Boc is tert-butoxycarbonyl protecting group (tert-butoxycarbonyl)
  • TBDMS/TBS is tert-butyldimethylsilyl protecting group ( tert-butyldimethylsilyl).
  • Fmoc-Val-OSu 100g, 229mmol was dissolved in 500ml of tetrahydrofuran, N ⁇ -(tert-butoxycarbonyl)-L-lysine (59.3g, 241mmol) and sodium bicarbonate (20.21g, 241mmol) were added respectively in 500 ml of aqueous solution.
  • the reaction solution was stirred at room temperature for 48 hours, and the detection reaction was completed.
  • the pH of the reaction solution was adjusted to about 6 with 1N dilute hydrochloric acid, and 500 ml of ethyl acetate was added for extraction.
  • the organic phase was separated, washed once with water and saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness.
  • compound (A-5) is the same as that of compound (A-4), except that the acetylated-10-polysarcosine (Ac-Sar10-COOH) in the last step is replaced by acetylated-4-polysarcosine Amino acid (Ac-Sar4-COOH), the product (A-5) is a beige amorphous powder after several steps of reaction.
  • LC-MS (ESI, m/z) theoretical: 1330.60, found: 1331.61 (M+H).
  • the intermediate compound 15-5 (150 mg, 0.131 mmol) was dissolved in 1 ml of anhydrous DMF, and acetylated-10 polysarcosine (Ac-Sar10-COOH) (101 mg, 0.131 mmol), DIPEA (114 ⁇ l, 0.654 mmol) and HATU (74.6 mg, 0.196 mmol), stirred at room temperature overnight, the solvent was removed under reduced pressure to obtain the final product, which was then prepared and purified by reverse-phase HPLC to obtain compound A-15 (107 mg, 0.059 mmol, yield 45.4%).
  • LC-MS (ESI, m/z) theoretical: 1799.74, found: 1800.77 (M+H).
  • the intermediate compound 15-5 (300 mg, 0.268 mmol) was dissolved in 5 ml of anhydrous DMF, and acetylated-10 polysarcosine (Ac-Sar10-COOH) (206 mg, 0.268 mmol), DIPEA (94 ⁇ l, 0.536 mmol) and HATU (122 mg, 0.321 mmol), stirred overnight at room temperature, and removed the solvent under reduced pressure to obtain the final product, which was then prepared and purified by reverse-phase HPLC to obtain compound A-18 (170 mg, 36% yield).
  • LC-MS (ESI, m/z) theoretical: 1757.78, found: 1758.82 (M+H).
  • Mc-Val-Cit-OH (purchased from Shanghai Haoyuan Chemical) reacts with bis(4-nitrophenyl) carbonate to obtain activated nitroester and further reacts with ixatecan methanesulfonic acid Salt (purchased from Shanghai Haoyuan) was reacted to obtain the reference compound MC-Val-Cit-PAB-DX8951.
  • the antibody or antigen-binding fragment eg, a stock solution of Trastuzumab targeting HER2 (heavy chain sequence is shown in SEQ ID NO: 3, light chain sequence is shown in SEQ ID NO: 4) and/or, for example, targeting Trop-2.
  • hRS7 mAb (Sacituzumab) in 50 mM potassium dihydrogen phosphate-sodium hydroxide (KH 2 PO4-NaOH)/150 mM sodium chloride (NaCl)/1 mM diethyltriaminepentaacetic acid (DTPA), pH 7 reaction buffer Dilute to 2 mg/mL, add tris(2-carboxyethyl)phosphine hydrochloride (TCEP) in an excess molar ratio of 6.0 times, and stir the reaction solution at 35° C. for 2.5 hours.
  • KH 2 PO4-NaOH potassium dihydrogen phosphate-sodium hydroxide
  • NaCl sodium chloride
  • DTPA diethyltriaminepentaacetic acid
  • pH 7 reaction buffer Dilute to 2 mg/mL, add tris(2-carboxyethyl)phosphine hydrochloride (TCEP) in an excess molar ratio of 6.0 times, and stir the reaction solution at 35°
  • the above reaction solution was cooled to 8°C, an appropriate amount of dimethylacetamide (DMA) was added without purification, and then 6-15 times excess molar ratio of reference substance drug molecules or drug linker conjugates A1 to A29 ( 10 mg/ml pre-dissolved in DMA) to ensure that the volume ratio of DMA in the reaction system does not exceed 20%, and the coupling was performed by stirring at 37° C. for 3 hours.
  • DMA dimethylacetamide
  • the coupling reaction mixture was purified by gel filtration of pH 6.0 histidine-acetic acid/sucrose using a desalting column, and the peak samples were collected according to the UV280 absorption value. It was then sterilized through a 0.15 micron pore size filter and stored at -60°C.
  • a 12-fold excess of compound (A-4) was conjugated with the reduced Trastuzumab monoclonal antibody to obtain antibody conjugate 4 with a drug loading (DAR) of about 7 to 8.
  • DAR drug loading
  • Size exclusion chromatography ( SEC-HPLC) to analyze the content of the polymer, and the analytical spectrum is shown in Figure 5.
  • the proportion of aggregate 1 is about 0.3%.
  • the drug loading (DAR) was about 7-8 by hydrophobic interaction high performance liquid chromatography (HIC-HPLC) analysis, and the analytical spectrum is shown in Figure 6.
  • the hRS7 single-antigen solution targeting Trop-2 was dialyzed to 50mM sodium dihydrogen phosphate-disodium hydrogen phosphate (NaH 2 PO 4 -Na 2 HPO 4 )/50mM sodium chloride (NaCl), pH7.0 buffer middle. After measuring the concentration of mAb in the solution, dilute the antibody to 5 mg/mL with the above buffer. The reaction tube was placed in an ice bath to cool down for 10 minutes. Tris(2-carboxyethyl)phosphine hydrochloride (TCEP) was added in a molar ratio of 2.5 times, and the reaction solution was stirred at 4°C overnight.
  • TCEP Tris(2-carboxyethyl)phosphine hydrochloride
  • DMA dimethylacetamide
  • a 6.0-fold excess molar ratio of the reference substance drug molecule or drug linker conjugate A-11 (10mM pre-dissolved in DMA) ensure that the volume ratio of DMA in the reaction system does not exceed 10%, and stir at 4 °C for 2 hours for coupling.
  • a 4.0-fold molar ratio of Cysteine (Cysteine) small molecule was added to the reaction solution to consume the excess drug linker conjugate A-11, and the reaction was stirred at 4°C for 30 minutes for quenching.
  • the coupling reaction mixture was purified by filtration with histidine-acetic acid/sodium chloride pH 5.5 using a desalting column, and the filtrated sample was collected.
  • One-tenth volume of activated carbon-histidine-acetic acid/sodium chloride suspension (300 mg/mL) was added to the sample, and stirred at room temperature for 2 hours to fully absorb free small drug molecules. It was then sterilized through a 0.22 micron pore size filter and stored at -80°C.
  • the content of the polymer was analyzed by size exclusion chromatography (SEC-HPLC), and the analysis pattern was shown in Figure 25.
  • the purity of the antibody conjugate 13 monomer was 98.95%.
  • the conjugated drug concentration in the antibody-drug conjugate can be calculated by measuring the UV absorbance of the antibody-drug conjugate aqueous solution at two wavelengths of 280 nm and 370 nm, and then calculated as follows.
  • the total absorbance at any given wavelength is equal to the sum of the absorbances of all light-absorbing chemicals in the system (absorption additivity). Therefore, based on the assumption that the molar absorption coefficient of the antibody and the drug does not change before and after conjugation of the antibody and the drug, the antibody concentration and the drug concentration in the antibody-drug conjugate are represented by the following formula.
  • a 280 represents the absorbance of the antibody-drug conjugate solution at 280 nm
  • a 370 represents the absorbance of the antibody-drug conjugate solution at 370 nm
  • AD,280 represents the absorbance of the drug-linker conjugate at 280 nm
  • AD,370 represents the absorbance of the drug-linker conjugate at 370 nm
  • a A,280 represents the absorbance of the antibody at 280 nm
  • a A,370 represents the absorbance of the antibody at 370 nm.
  • ⁇ D,280 represents the molar absorption coefficient of the drug-linker conjugate at 280 nm
  • ⁇ D,370 represents the molar absorption coefficient of the drug-linker conjugate at 370 nm
  • ⁇ A,280 represents the molar absorption of the antibody at 280 nm
  • the coefficient, ⁇ A, 370 represents the molar absorption coefficient of the antibody at 370 nm.
  • CD represents the concentration of the drug linker conjugate in the antibody-drug conjugate solution
  • CA represents the concentration of the antibody in the antibody-drug conjugate solution.
  • ⁇ A,280 can be estimated from the amino acid sequence of the antibody by known computational methods (Protein Science, 1995, vol. 4, 2411-2423), and ⁇ A,370 is generally zero.
  • ⁇ D,280 and ⁇ D,370 are obtained according to the Lambert-Beer law by the absorbance of the solution of the drug-linker conjugate at a certain molar concentration.
  • the values of A 280 and A 370 are measured by a microplate reader or UV spectrophotometer, and the values of the above 4 molar absorption coefficients are put into the simultaneous equations (1) and (2), and C A and C can be calculated value of D.
  • the average number of conjugated drug molecules in each antibody molecule DAR C D /C A .
  • the hRS7 single-antigen solution targeting Trop-2 was dialyzed to 50mM sodium dihydrogen phosphate-disodium hydrogen phosphate (NaH 2 PO 4 -Na 2 HPO 4 )/50mM sodium chloride (NaCl), pH7.0 buffer middle. After measuring the concentration of mAb in the solution, dilute the antibody to 5 mg/mL with the above buffer. The reaction tube was placed in an ice bath to cool down for 10 minutes. Tris(2-carboxyethyl)phosphine hydrochloride (TCEP) was added in a molar ratio of 2.5 times, and the reaction solution was stirred at 4°C overnight.
  • TCEP Tris(2-carboxyethyl)phosphine hydrochloride
  • DMA dimethylacetamide
  • the coupling reaction mixture was purified by filtration with histidine-acetic acid/sodium chloride pH 5.5 using a desalting column, and the filtrated sample was collected.
  • One-tenth volume of activated carbon-histidine-acetic acid/sodium chloride suspension 300 mg/mL was added to the sample, and stirred at room temperature for 2 hours to fully absorb free small drug molecules. It was then sterilized through a 0.22 micron pore size filter and stored at -80°C.
  • the content of the polymer was analyzed by size exclusion chromatography (SEC-HPLC), and the analysis pattern was shown in Figure 26.
  • the purity of the antibody conjugate 14 monomer was 98.20%.
  • the hRS7 single-antigen solution targeting Trop-2 was dialyzed to 50mM sodium dihydrogen phosphate-disodium hydrogen phosphate (NaH 2 PO 4 -Na 2 HPO 4 )/50mM sodium chloride (NaCl), pH7.0 buffer middle. After measuring the concentration of mAb in the solution, dilute the antibody to 5 mg/mL with the above buffer. The reaction tube was placed in an ice bath to cool down for 10 minutes. Tris(2-carboxyethyl)phosphine hydrochloride (TCEP) was added in a molar ratio of 2.5 times, and the reaction solution was stirred at 4°C overnight.
  • TCEP Tris(2-carboxyethyl)phosphine hydrochloride
  • DMA dimethylacetamide
  • the coupling reaction mixture was purified by filtration with histidine-acetic acid/sodium chloride pH 5.5 using a desalting column, and the filtrated sample was collected.
  • One-tenth volume of activated carbon-histidine-acetic acid/sodium chloride suspension 300 mg/mL was added to the sample, and stirred at room temperature for 2 hours to fully absorb free small drug molecules. It was then sterilized through a 0.22 micron pore size filter and stored at -80°C.
  • the content of the polymer was analyzed by size exclusion chromatography (SEC-HPLC), and the analysis pattern was shown in Figure 27.
  • the purity of the antibody conjugate 15 monomer was 96.36%.
  • ADC drugs based on camptothecin-like topoisomerase inhibitors such as Sacituzumab govitecan (Trodelvy) and Trastuzumab Deruxtecan (Enhertu) usually have a higher drug-to-antibody loading ratio (DAR), due to the aromatic concentration of camptothecin-like molecules.
  • DAR drug-to-antibody loading ratio
  • the ring-like structure has strong hydrophobicity, and the stability of such ADC molecules must be affected to a certain extent.
  • the ADC prepared by the present invention introduces a polysarcosine peptide chain at a specific position, which has better "barrier effect" and may have better stability. We designed an antibody stability accelerated test to verify this hypothesis.
  • Molecules compared include:
  • the mobile phase of HPLC was 200 mM phosphate buffer (pH 7.0) + 150 mM KCl + 15% IPA, the column temperature was 25 °C, the injection volume was 7 ⁇ L, the flow rate was 0.75 ml/min, and the UV detection wavelengths were 280 nm and 370 nm.
  • Human lung squamous cell carcinoma cells NCI-H2170 and LK-2, and human breast cancer cell MDA-MB-231 were selected as the cell lines for in vitro activity detection in this experiment, and the surface TROP2 antigen of the above human tumor cells was determined by flow cytometry (FACS).
  • the expression level (expressed by rMFI), and the effect of antibody conjugate 13, DS-1062a (control drug, prepared according to the method described in US11173213B2), hIgG-deruxtecan conjugate, and hIgG-A-11 conjugate on cells was observed at the same time. The dose-response situation of killing.
  • the final concentration of the antibody conjugated drug after adding the sample was set at 500nM as the initial concentration, and 500-0.1nM was designed in a series of 9 concentrations (5-fold ratio dilution), and the killing (or inhibition) changes were observed for 120 hours, and chemiluminescence staining (Luminescent Cell Viability Assay), IC 50 was calculated after reading the fluorescence data.
  • both antibody conjugate 13 and DS-1062a can significantly inhibit the proliferation of tumor cells, which are significantly stronger than the negative and positive drugs IgG-deruxtecan conjugate and IgG A-1 conjugate.
  • the inventive antibody conjugate 13 is significantly better than the clinical drug DS-1062a.
  • the ADCs prepared from the compounds of the present application all show good in vitro anti-tumor activity, especially for tumor cells with low expression of related antigens.
  • the antibody conjugates of the present application have more obvious advantages.
  • the efficacy of the combinations of the invention can be measured in vivo, ie in rodents implanted with cancer cell allografts or xenografts, and treated with the combination.
  • Test mice are treated with drug or control and monitored for several weeks or longer to measure time to tumor doubling, log cell killing, and tumor inhibition.
  • Human lung squamous cell carcinoma cell NCI-H2170 (ATCC) was suspended in physiological saline, 4 ⁇ 10 7 cells were subcutaneously transplanted into the right flank of female nude mice, and grouping was performed at random on the sixth day. Taking the grouping day as the 0th day, on the 0th day, the antibody conjugate 13, DS-1062a (positive control drug) and hIgG-A-11 conjugate were respectively administered to the tail vein at a dose of 5 mg/kg as the negative control group.
  • the ADCs prepared from the compounds of the present application all showed a certain anti-tumor activity in vivo, and could show significantly stronger anti-tumor activity compared with the control sample.
  • the tumor-bearing mice tolerated the above drugs well, and no symptoms such as weight loss occurred.

Abstract

The present application relates to a preparation method for an antibody-drug conjugate and an application, and in particular, to a compound, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, a preparation method for the compound and related antibody-drug conjugates, and a use of same in the preparation of drugs for treating cancer.

Description

抗体偶联药物的制备方法及应用Preparation method and application of antibody conjugated drug 技术领域technical field
本申请涉及生物医药领域，具体的涉及一种抗体偶联药物的制备方法及应用。The present application relates to the field of biomedicine, in particular to a preparation method and application of an antibody conjugated drug.
背景技术Background technique
作为新型的靶向治疗药物，抗体药物偶联物(Antibody Drug Conjugate,ADC)通过适当的载体将药物靶向输送到疾患部位。然而，受制于抗体及高活性细胞毒性药物技术的制约，有限数量的抗体药物偶联物获得批准上市，抗体药物偶联物在肿瘤治疗领域的应用急需快速发展。As a new type of targeted therapy drug, Antibody Drug Conjugate (ADC) is used to deliver the drug to the diseased site through an appropriate carrier. However, limited by the technology of antibodies and highly active cytotoxic drugs, a limited number of antibody-drug conjugates have been approved for marketing, and the application of antibody-drug conjugates in the field of tumor therapy is in urgent need of rapid development.
细胞毒物对ADC药物发挥作用是必不可少的，喜树碱类药物应用前景巨大，上市ADC药物Trodelvy和Enhertu即为分别采用喜树碱类药物SN38和DX-8951f作为弹头分子。然而通过喜树碱类药物最终制备得到的ADC都极大改变了单抗的属性，稳定性和半衰期都有较大程度衰减。虽然在DESTINY-Breast03临床研究中，Enhertu确认的客观缓解率(ORR)达到79.7％，然而根据DS8201-A-J101临床研究显示对于HER2低表达乳腺癌，Enhertu确认的客观缓解率(ORR)为37.0％，治疗有效率明显低于高表达或中表达的乳腺癌。此外，ADC药物在毒副作用上出现了新的症状，如药物Enhertu出现了肺炎和间质性肺病等副作用。WO2020233174A1公开了一类含有缬氨酸-瓜氨酸(Val-Cit)-PAB连接子的喜树碱类化合物，然而该分子无法与抗体偶联后获得质量合格的ADC分子(合格的ADC产品中聚体含量需要小于5％)。因此，本领域迫切需要提供更为合适的基于喜树碱类药物(如依沙替康、贝洛替康等)的抗体偶联药物，实现高效、简单、实用的化学制备与偶联，并且改善现有抗体偶联药物的药学性质、代谢性质、药效及安全性等，如提高ADC分子稳定性、提高治疗窗口等。Cytotoxics are essential for ADC drugs to play a role. Camptothecin drugs have great application prospects. The marketed ADC drugs Trodelvy and Enhertu use camptothecin drugs SN38 and DX-8951f as warhead molecules respectively. However, the ADCs finally prepared by camptothecins have greatly changed the properties of monoclonal antibodies, and the stability and half-life have been greatly attenuated. Although in the DESTINY-Breast03 clinical study, the objective response rate (ORR) confirmed by Enhertu reached 79.7%, according to the clinical study DS8201-A-J101, the objective response rate (ORR) confirmed by Enhertu was 37.0 for HER2-low expressing breast cancer %, the treatment efficiency was significantly lower than that of breast cancer with high or medium expression. In addition, ADC drugs have new symptoms of toxic side effects, such as pneumonia and interstitial lung disease with the drug Enhertu. WO2020233174A1 discloses a class of camptothecin compounds containing a valine-citrulline (Val-Cit)-PAB linker, however, this molecule cannot be coupled with an antibody to obtain a qualified ADC molecule (in qualified ADC products). The aggregate content needs to be less than 5%). Therefore, there is an urgent need in the art to provide more suitable antibody-drug conjugates based on camptothecins (such as ixatecan, belotecan, etc.) to achieve efficient, simple and practical chemical preparation and conjugation, and Improve the pharmaceutical properties, metabolic properties, efficacy and safety of the existing antibody-conjugated drugs, such as improving the stability of ADC molecules, improving the therapeutic window, etc.
发明内容SUMMARY OF THE INVENTION
本申请提供了一种抗体偶联药物、其中间体、制备方法及应用。本申请的抗体偶联药物，可以实现细胞毒性药物在ADC领域的广泛应用，治疗肿瘤疾病。本申请的主要技术效果在于：提供的新型连接子，可通过特定的化学方法将疏水性极强的抗肿瘤药物，如拓扑异构酶抑制剂依沙替康、贝洛替康等与抗体进行偶联，所获得的偶联物具有高亲水性和稳定性，相较于传统的ADC连接子，本申请提供的连接子在较高载药量时抗体偶联物不易产生聚体；相较于同类ADC药物，本申请释放后的毒素分子为拓扑异构酶抑制剂依沙替康、贝洛替康原型分子，通过测试表明该药物分子较同类ADC药物所释放的药物衍生物具有更好的生物 活性、安全性以及其它药物相关性质。因此，本申请可以提高药物体内半衰期以及药物在肿瘤组织的浓度，从而提到药物的抗肿瘤活性和/或提高整体药物治疗窗口。The present application provides an antibody conjugated drug, an intermediate thereof, a preparation method and an application thereof. The antibody conjugated drug of the present application can realize the wide application of cytotoxic drugs in the field of ADC to treat tumor diseases. The main technical effect of the present application is that the provided novel linker can combine highly hydrophobic anti-tumor drugs, such as topoisomerase inhibitors isanotecan, belotecan, etc., with antibodies through specific chemical methods. Conjugation, the obtained conjugate has high hydrophilicity and stability. Compared with the traditional ADC linker, the antibody conjugate provided by the application is not easy to produce aggregates when the drug load is higher; Compared with similar ADC drugs, the toxin molecules released in the present application are the prototype molecules of topoisomerase inhibitor ixatecan and belotecan. Tests show that the drug molecules have better properties than the drug derivatives released by similar ADC drugs. Good biological activity, safety and other drug-related properties. Therefore, the present application can improve the half-life of the drug in vivo and the concentration of the drug in the tumor tissue, thereby referring to the anti-tumor activity of the drug and/or improving the overall drug treatment window.
44.一方面，本申请提供了一种化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，其中所述化合物包含式(C-Trop-2)所示的结构：44. In one aspect, the application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or A pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the compound comprises the structure represented by the formula (C-Trop-2):
Figure PCTCN2022089726-appb-000001
Figure PCTCN2022089726-appb-000001
其中，Q 1包含连接体， where Q1 contains the linker,
L 1包含-L 1a-C(＝O)-， L 1 contains -L 1a -C(=O)-,
其中，L 1a选自以下组：任选取代的亚烷基、任选取代的聚乙二醇基、任选取代的亚烯基、任选取代的亚炔基、任选取代的亚脂环基、任选取代的亚脂杂环基、任选取代的亚芳基和任选取代的亚杂芳基； wherein, L 1a is selected from the group consisting of optionally substituted alkylene, optionally substituted polyethylene glycol, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted alicyclic groups, optionally substituted alicyclic heterocyclyl groups, optionally substituted arylene groups, and optionally substituted heteroarylene groups;
L 2包含任选取代的多肽残基， L2 comprises optionally substituted polypeptide residues,
L 3包含任选取代的间隔基团， L 3 comprises an optionally substituted spacer group,
其中，L 2和/或L 3包含任选取代的聚肌氨酸残基， wherein L 2 and/or L 3 comprise optionally substituted polysarcosine residues,
T包含药物单元，T contains the drug unit,
Ab为能够结合Trop-2的配体，m为1-8的数。Ab is a ligand capable of binding Trop-2, and m is a number from 1 to 8.
另一方面，本申请提供了一种化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，In another aspect, the application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a mixture thereof A pharmaceutically acceptable salt, prodrug or solvate,
其中，所述化合物包含选自以下组的结构：wherein the compound comprises a structure selected from the group consisting of:
Figure PCTCN2022089726-appb-000002
Figure PCTCN2022089726-appb-000002
Figure PCTCN2022089726-appb-000003
Figure PCTCN2022089726-appb-000003
Figure PCTCN2022089726-appb-000004
Figure PCTCN2022089726-appb-000004
Ab为能够结合Trop-2的配体，m为1-8的数。Ab is a ligand capable of binding Trop-2, and m is a number from 1 to 8.
另一方面，本申请提供了一种药物组合物，其含有本申请任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，以及任选地药学上可接受的载体。On the other hand, the application provides a pharmaceutical composition, which contains the compound described in any one of the application, or its tautomer, meso, racemate, enantiomer, A diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, and optionally a pharmaceutically acceptable carrier.
另一方面，本申请提供了含有本申请任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，和/或本申请所述的药物组合物在制备用于治疗和/或预防肿瘤的药物中的用途。On the other hand, the present application provides a compound containing any one of the present application, or its tautomer, meso, racemate, enantiomer, diastereomer, Use of the mixture form thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, and/or the pharmaceutical composition described in this application in the preparation of a medicament for treating and/or preventing tumors.
另一方面，本申请提供了一种抗体-药物偶联物，所述的抗体-药物偶联物通过本申请的化合物与抗体进行偶联形成的。In another aspect, the present application provides an antibody-drug conjugate, which is formed by coupling the compound of the present application with an antibody.
在一种实施方式中，本申请的偶联物共价连接有一个或多个药物组分。In one embodiment, the conjugates of the present application are covalently attached to one or more drug components.
在一种实施方式中，本申请的抗体和药物是通过共价方式(例如，可以通过分别共价连接于连接子上)进行偶联。In one embodiment, the antibody and drug of the present application are coupled covalently (eg, by covalent attachment to a linker, respectively).
另一方面，本申请提供一种抗体-药物偶联物的制备方法，通过所述抗体或抗体片断铰 链区的二硫链还原生成一对半胱氨酸残基，并通过所述半胱氨酸残基中巯基与本申请中化合物的连接基团，例如马来酰亚胺基发生取代反应，进而将本申请的化合物连接于所述抗体或抗体片段的半胱氨酸巯基上，获得抗体-药物偶联物，药物抗体偶联率DAR(例如本申请中的m)根据反应条件进行控制，例如常见为2～8之间。In another aspect, the present application provides a method for preparing an antibody-drug conjugate, wherein a pair of cysteine residues is generated by reduction of the disulfide chain in the hinge region of the antibody or antibody fragment, and the cysteine The sulfhydryl group in the acid residue undergoes a substitution reaction with the connecting group of the compound in the present application, such as maleimide group, and then the compound of the present application is connected to the cysteine sulfhydryl group of the antibody or antibody fragment to obtain an antibody -Drug conjugate, the drug-antibody conjugation rate DAR (for example, m in this application) is controlled according to the reaction conditions, for example, it is usually between 2 and 8.
另一方面，本申请中m表示细胞毒性药物分子与Ab的摩尔比(又称DAR，即药物抗体偶联比率)，m可为整数或小数，可以理解为是：单个单克隆抗体分子与细胞毒性药物偶联后得到的抗体偶联药物中的药物分子与单克隆抗体分子的摩尔比的平均值，一般可以采用疏水层析(Hydrophobic-Interaction Chromatography，HIC)，反相高效液相色谱(Reverse phase HPLC,RP-HPLC)、聚丙烯酰胺-SDS凝胶电泳(SDS PAGE，electrophoresis)，液相质谱(liquid chromatograph-mass spectrometer，LC-MS)、紫外/可见光谱法(UV/Vis)等方式测定得到。On the other hand, in this application, m represents the molar ratio of cytotoxic drug molecules to Ab (also known as DAR, that is, drug-antibody coupling ratio), and m can be an integer or a decimal number, which can be understood as: a single monoclonal antibody molecule and cell The average molar ratio of drug molecules to monoclonal antibody molecules in the antibody-drug conjugate obtained after toxic drug conjugation can generally be determined by hydrophobic chromatography (Hydrophobic-Interaction Chromatography, HIC), reversed-phase high-performance liquid chromatography (Reverse high-performance liquid chromatography) phase HPLC, RP-HPLC), polyacrylamide-SDS gel electrophoresis (SDS PAGE, electrophoresis), liquid chromatograph-mass spectrometer (LC-MS), ultraviolet/visible spectroscopy (UV/Vis), etc. measured.
另一方面，本申请提供一种制备本申请的化合物的方法，其包含以下步骤：On the other hand, the application provides a method for preparing the compound of the application, comprising the steps of:
使带有氨基保护基团N1的氨基酸活性酯与氨基酸接触，获得中间体M1；在缩合剂如EEDQ(2-乙氧基-1-乙氧碳酰基-1,2-二氢喹啉)存在下，使所述中间体M1与取代/非取代对氨基苄醇接触，获得中间体M2；脱除所述中间体M2的所述N1，获得中间体M3；使所述中间体M3与含有马亚酰亚胺基团的化合物接触，获得中间体M4；使所述中间体M4与双(4-硝基苯基)碳酸酯接触，获得中间体M5；使所述中间体M5与药物单元接触。Contact an amino acid active ester with an amino protecting group N1 with an amino acid to obtain an intermediate M1; in the presence of a condensing agent such as EEDQ (2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline) Next, the intermediate M1 is contacted with substituted/unsubstituted p-aminobenzyl alcohol to obtain the intermediate M2; the N1 of the intermediate M2 is removed to obtain the intermediate M3; contacting a compound of an imide group to obtain an intermediate M4; contacting the intermediate M4 with bis(4-nitrophenyl) carbonate to obtain an intermediate M5; contacting the intermediate M5 with a drug unit .
在一种实施方式中，所述N1包含芴基甲氧基羰基。In one embodiment, the N1 comprises fluorenylmethoxycarbonyl.
在一种实施方式中，当制备所述L 2被包含聚肌氨酸残基的结构取代的所述化合物时，使包含氨基保护基团N2的中间体与三氟乙酸接触后，再与乙酰化的多聚肌氨酸接触。 In one embodiment, when preparing the compound in which the L2 is substituted by a structure comprising a polysarcosine residue, the intermediate comprising the amino protecting group N2 is contacted with trifluoroacetic acid, followed by contact with acetyl polysarcosine contacts.
在一种实施方式中，所述N2包含叔丁氧羰基。In one embodiment, the N2 comprises tert-butoxycarbonyl.
另一方面，本申请提供一种制备本申请的化合物的方法，其包含以下步骤：在适于在配体和化合物之间形成键的条件下，使所述配体与本申请的化合物接触。In another aspect, the application provides a method of preparing a compound of the application comprising the steps of contacting the ligand with the compound of the application under conditions suitable for forming a bond between the ligand and the compound.
在一种实施方式中，所述配体与本申请的化合物在缓冲液与有机溶剂混合液中接触。In one embodiment, the ligand is contacted with the compound of the present application in a mixture of buffer and organic solvent.
在一种实施方式中，所述配体与本申请的化合物在约0至约37℃下接触。In one embodiment, the ligand is contacted with a compound of the present application at about 0 to about 37°C.
在一种实施方式中，在使所述配体与本申请的化合物接触之前包含以下步骤：将所述配体与还原试剂在缓冲液中反应，得到经还原后的所述配体。In one embodiment, the following step is included before contacting the ligand with the compound of the present application: reacting the ligand with a reducing agent in a buffer to obtain the reduced ligand.
在一种实施方式中，在得到经还原后的所述配体之后，且在使所述配体与本申请的化合物接触之前包含以下步骤：去除所述还原试剂。In one embodiment, after obtaining the reduced ligand, and before contacting the ligand with the compound of the present application, the step of removing the reducing agent is included.
在一种实施方式中，所述去除所述还原试剂包含将反应产物进行过脱盐柱和/或超滤。In one embodiment, the removing the reducing agent comprises subjecting the reaction product to a desalting column and/or ultrafiltration.
在一种实施方式中，所述还原试剂选自以下组：三(2-羧乙基)膦盐酸盐(TCEP)、beta-巯 基乙醇、beta-巯基乙胺盐酸盐和二硫苏糖醇(DTT)。In one embodiment, the reducing agent is selected from the group consisting of tris(2-carboxyethyl)phosphine hydrochloride (TCEP), beta-mercaptoethanol, beta-mercaptoethylamine hydrochloride, and dithiothreose Alcohol (DTT).
在一种实施方式中，所述缓冲液选自以下组：磷酸二氢钾-氢氧化钠(KH 2PO 4-NaOH)/氯化钠(NaCl)/二乙基三胺五乙酸(DTPA)缓冲液、磷酸氢二钠-柠檬酸/氯化钠(NaCl)/二乙基三胺五乙酸(DTPA)、硼酸-硼砂/氯化钠(NaCl)/二乙基三胺五乙酸(DTPA)、组氨酸-氢氧化钠/氯化钠(NaCl)/二乙基三胺五乙酸(DTPA)和PBS/二乙基三胺五乙酸(DTPA)。 In one embodiment, the buffer is selected from the group consisting of potassium dihydrogen phosphate-sodium hydroxide (KH 2 PO 4 -NaOH)/sodium chloride (NaCl)/diethyltriaminepentaacetic acid (DTPA) Buffer, disodium hydrogen phosphate-citric acid/sodium chloride (NaCl)/diethyltriaminepentaacetic acid (DTPA), boric acid-borax/sodium chloride (NaCl)/diethyltriaminepentaacetic acid (DTPA) , histidine-sodium hydroxide/sodium chloride (NaCl)/diethyltriaminepentaacetic acid (DTPA) and PBS/diethyltriaminepentaacetic acid (DTPA).
在一种实施方式中，所述有机溶剂选自以下组：乙腈(ACN)、二甲基甲酰胺(DMF)、二甲基乙酰胺(DMA)和二甲基亚砜(DMSO)。In one embodiment, the organic solvent is selected from the group consisting of acetonitrile (ACN), dimethylformamide (DMF), dimethylacetamide (DMA) and dimethylsulfoxide (DMSO).
在一种实施方式中，所述有机溶剂在所述缓冲液与有机溶剂混合液中的体积占比不超过30％。In one embodiment, the volume ratio of the organic solvent in the buffer solution and the organic solvent mixture does not exceed 30%.
本申请发现本申请中的抗体-药物偶联物，与传统抗体-药物偶联物相比，由于依沙替康、贝洛替康类药物疏水性过大，而聚肌氨酸的引入可以大大增加偶联物的亲水性，从而从整体上使得所获得抗体-药物偶联物更加稳定性，不易聚合。此外，本申请提供了氨基甲酸酯，且能在酶切后发生快速的1,6-消除从而释放药物，具有更好的体外与体内的稳定性与生物活性。基于上述发现，完成了本申请。This application finds that the antibody-drug conjugates in this application, compared with traditional antibody-drug conjugates, due to the large hydrophobicity of ixatecan and belotecan drugs, the introduction of polysarcosine can The hydrophilicity of the conjugate is greatly increased, thereby making the obtained antibody-drug conjugate more stable and less prone to polymerization as a whole. In addition, the present application provides carbamate, which can undergo rapid 1,6-elimination after enzymatic cleavage to release the drug, and has better stability and biological activity in vitro and in vivo. Based on the above findings, the present application has been completed.
本领域技术人员能够从下文的详细描述中容易地洞察到本申请的其它方面和优势。下文的详细描述中仅显示和描述了本申请的示例性实施方式。如本领域技术人员将认识到的，本申请的内容使得本领域技术人员能够对所公开的具体实施方式进行改动而不脱离本申请所涉及发明的精神和范围。相应地，本申请的附图和说明书中的描述仅仅是示例性的，而非为限制性的。Other aspects and advantages of the present application can be readily appreciated by those skilled in the art from the following detailed description. Only exemplary embodiments of the present application are shown and described in the following detailed description. As those skilled in the art will recognize, the content of this application enables those skilled in the art to make changes to the specific embodiments disclosed without departing from the spirit and scope of the invention to which this application relates. Accordingly, the drawings and descriptions in the specification of the present application are only exemplary and not restrictive.
附图说明Description of drawings
本申请所涉及的发明的具体特征如所附权利要求书所显示。通过参考下文中详细描述的示例性实施方式和附图能够更好地理解本申请所涉及发明的特点和优势。对附图简要说明如下：The invention to which this application relates is set forth with particularity characteristic of the appended claims. The features and advantages of the inventions involved in this application can be better understood by reference to the exemplary embodiments described in detail hereinafter and the accompanying drawings. A brief description of the drawings is as follows:
图1显示的是本申请抗体偶联物1的尺寸排阻层析(SEC-HPLC)色谱图；Figure 1 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 1 of the present application;
图2显示的是本申请抗体偶联物2的尺寸排阻层析(SEC-HPLC)色谱图；Figure 2 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 2 of the present application;
图3显示的是本申请抗体偶联物2的疏水作用高效液相色谱图(HIC-HPLC)；Figure 3 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 2 of the present application;
图4显示的是本申请抗体偶联物3的尺寸排阻层析(SEC-HPLC)色谱图；Figure 4 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 3 of the present application;
图5显示的是本申请抗体偶联物4的尺寸排阻层析(SEC-HPLC)色谱图；Figure 5 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 4 of the present application;
图6显示的是本申请抗体偶联物4的疏水作用高效液相色谱图(HIC-HPLC)；Figure 6 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 4 of the present application;
图7显示的是本申请抗体偶联物5的尺寸排阻层析(SEC-HPLC)色谱图；Figure 7 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 5 of the present application;
图8显示的是本申请抗体偶联物5的疏水作用高效液相色谱图(HIC-HPLC)；Figure 8 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 5 of the present application;
图9显示的是本申请抗体偶联物6的尺寸排阻层析(SEC-HPLC)色谱图；Figure 9 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 6 of the present application;
图10显示的是本申请抗体偶联物6的疏水作用高效液相色谱图(HIC-HPLC)；Figure 10 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 6 of the present application;
图11显示的是本申请抗体偶联物7的尺寸排阻层析(SEC-HPLC)色谱图；Figure 11 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 7 of the present application;
图12显示的是本申请抗体偶联物7的疏水作用高效液相色谱图(HIC-HPLC)；Figure 12 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 7 of the present application;
图13显示的是本申请抗体偶联物8的尺寸排阻层析(SEC-HPLC)色谱图；Figure 13 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 8 of the present application;
图14显示的是本申请抗体偶联物8的疏水作用高效液相色谱图(HIC-HPLC)；Figure 14 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 8 of the present application;
图15显示的是本申请抗体偶联物9的尺寸排阻层析(SEC-HPLC)色谱图；Figure 15 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 9 of the present application;
图16显示的是本申请抗体偶联物9的疏水作用高效液相色谱图(HIC-HPLC)；Figure 16 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 9 of the present application;
图17显示的是本申请抗体偶联物10的尺寸排阻层析(SEC-HPLC)色谱图；Figure 17 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 10 of the present application;
图18显示的是本申请抗体偶联物10的疏水作用高效液相色谱图(HIC-HPLC)；Figure 18 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 10 of the present application;
图19显示的是本申请抗体偶联物11的尺寸排阻层析(SEC-HPLC)色谱图；Figure 19 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 11 of the present application;
图20显示的是本申请抗体偶联物11的疏水作用高效液相色谱图(HIC-HPLC)；Figure 20 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 11 of the present application;
图21显示的是本申请抗体偶联物12的尺寸排阻层析(SEC-HPLC)色谱图；Figure 21 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 12 of the present application;
图22显示的是本申请抗体偶联物12的疏水作用高效液相色谱图(HIC-HPLC)；Figure 22 shows the hydrophobic interaction high performance liquid chromatogram (HIC-HPLC) of the antibody conjugate 12 of the present application;
图23显示的是本申请部分抗体偶联物的加速稳定性实验浓度变化图；Figure 23 shows the concentration change diagram of the accelerated stability experiment of some antibody conjugates of the present application;
图24显示的是本申请部分抗体偶联物的加速稳定性实验聚体增加变化图；Figure 24 shows a graph showing the increase of aggregates in the accelerated stability experiment of some antibody conjugates of the present application;
图25显示的是本申请抗体偶联物13的尺寸排阻层析(SEC-HPLC)色谱图；Figure 25 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 13 of the present application;
图26显示的是本申请抗体偶联物14的尺寸排阻层析(SEC-HPLC)色谱图；Figure 26 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 14 of the present application;
图27显示的是本申请抗体偶联物15的尺寸排阻层析(SEC-HPLC)色谱图；Figure 27 shows the size exclusion chromatography (SEC-HPLC) chromatogram of the antibody conjugate 15 of the present application;
图28显示的是本申请部分抗体偶联物在人肺鳞癌细胞NCI-H2170体内药效结果对比图；Figure 28 shows a comparison chart of the efficacy results of some antibody conjugates of the present application in human lung squamous cell carcinoma NCI-H2170;
图29显示的是本申请部分抗体偶联物在人肺鳞癌细胞NCI-H2170体内药效实验中小鼠体重变化图。Figure 29 is a graph showing the changes in the body weight of mice in the in vivo efficacy experiment of partial antibody conjugates of the present application in human lung squamous cell carcinoma cells NCI-H2170.
具体实施方式Detailed ways
以下由特定的具体实施例说明本申请发明的实施方式，熟悉此技术的人士可由本说明书所公开的内容容易地了解本申请发明的其他优点及效果。The embodiments of the invention of the present application are described below with specific specific examples, and those skilled in the art can easily understand other advantages and effects of the invention of the present application from the contents disclosed in this specification.
术语定义Definition of Terms
在本申请中，术语“配体”通常指能识别和结合目标细胞相关的抗原或受体的大分子化合物。配体的作用可以是将药物呈递给与配体结合的目标细胞群，这些配体包括但不限于蛋白 类激素、凝集素、生长因子、抗体或其他能与细胞、受体和/或抗原结合的分子。在本申请中，配体可以表示为Ab，配体抗原通过配体上的杂原子与连接单元形成连接键，可以为抗体或其抗原结合片段，所述抗体可以选自嵌合抗体、人源化抗体、全人抗体或鼠源抗体；所述抗体可以是单克隆抗体。例如所述抗体可以是，靶向选自以下组的靶点的抗体或其抗原结合片段：HER2。In this application, the term "ligand" generally refers to a macromolecular compound capable of recognizing and binding to an antigen or receptor associated with a target cell. The role of the ligand can be to present the drug to the target cell population that binds to the ligand, including but not limited to protein hormones, lectins, growth factors, antibodies, or others that can bind to cells, receptors and/or antigens molecule. In this application, the ligand can be represented as Ab, and the ligand antigen forms a bond with the connecting unit through the heteroatom on the ligand, which can be an antibody or an antigen-binding fragment thereof, and the antibody can be selected from chimeric antibodies, human-derived antibody, fully human, or murine; the antibody may be a monoclonal antibody. For example, the antibody may be an antibody or antigen-binding fragment thereof targeting a target selected from the group consisting of: HER2.
在本申请中，术语“烷基”通常是指烷除去氢原子所衍生的残基。烷基可以是取代的或非取代的，替代或者非替代的。术语“烷基”通常指饱和的直链或支链脂肪族烃基，其具有从母体烷的相同碳原子或两个不同的碳原子上除去氢原子所衍生的残基，其可以为包含1至20个碳原子的直链或支链基团，例如含有1至12个碳原子，例如含有1至6个碳原子的链烷基。烷基的非限制性实例包括但不限于甲基、乙基、丙基、丙基、丁基等。烷基可以是取代的或非取代的，替代或者非替代的，例如当被取代时，取代基可以在任何可使用的连接点上被取代，所述取代基可以独立地任选选自烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基和氧代基中的一个或多个取代基所取代，例如可以是氢、氕、氘、氚、卤素、-NO 2、-CN、-OH、-SH、-NH 2、-C(O)H、-CO 2H、-C(O)C(O)H、-C(O)CH 2C(O)H、-S(O)H、-S(O) 2H、-C(O)NH 2、-SO 2NH 2、-OC(O)H、-N(H)SO 2H或C 1-6脂肪族基团。 In this application, the term "alkyl" generally refers to a residue derived from an alkane by removal of a hydrogen atom. Alkyl groups can be substituted or unsubstituted, substituted or unsubstituted. The term "alkyl" generally refers to a saturated straight-chain or branched aliphatic hydrocarbon group having a residue derived by removing a hydrogen atom from the same carbon atom or two different carbon atoms of the parent alkane, which may be a group containing 1 to A straight or branched chain group of 20 carbon atoms, eg 1 to 12 carbon atoms, eg, an alkane alkyl group containing 1 to 6 carbon atoms. Non-limiting examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, propyl, butyl, and the like. Alkyl groups may be substituted or unsubstituted, substituted or non-substituted, for example when substituted, substituents may be substituted at any available point of attachment, and the substituents may be independently optionally selected from alkyl groups , alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy , heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and one or more substituents in oxo, such as hydrogen, protium, deuterium, tritium, halogen, -NO 2 , - CN, -OH, -SH, -NH2 , -C(O)H, -CO2H , -C(O)C(O)H, -C(O) CH2C (O)H, -S (O)H, -S(O)2H, -C (O) NH2 , -SO2NH2 , -OC ( O)H, -N(H) SO2H or C1-6 aliphatic group.
在本申请中，术语“亚烷基”通常指饱和的直链或支链脂肪族烃基，其具有2个从母体烷的相同碳原子或两个不同的碳原子上除去两个氢原子所衍生的残基，其可以为包含1至20个碳原子的直链或支链基团，例如，术语“亚甲基”可以是指1个碳原子的基团除去两个氢原子所衍生的残基。亚甲基可以是取代的或非取代的，替代或者非替代的；例如含有1至12个碳原子，例如含有1至6个碳原子的亚烷基。亚烷基的非限制性实例包括但不限于亚甲基(-CH 2-)、1,1-亚乙基(-CH(CH 3)-)、1,2-亚乙基(-CH 2CH 2)-、1,1-亚丙基(-CH(CH 2CH 3)-)、1,2-亚丙基(-CH 2CH(CH 3)-)、1,3-亚丙基(-CH 2CH 2CH 2-)、1,4-亚丁基(-CH 2CH 2CH 2CH 2-)和1,5-亚丁基(-CH 2CH 2CH 2CH 2CH 2-)等。亚烷基可以是取代的或非取代的，替代或者非替代的，例如当被取代时，取代基可以在任何可使用的连接点上被取代，所述取代基可以独立地任选选自烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基和氧代基中的一个或多个取代基所取代，例如可以是氢、氕、氘、氚、卤素、-NO 2、-CN、-OH、-SH、-NH 2、-C(O)H、-CO2H、-C(O)C(O)H、-C(O)CH 2C(O)H、-S(O)H、-S(O) 2H、-C(O)NH 2、-SO 2NH 2、-OC(O)H、-N(H)SO 2H或C 1-6脂肪族基团。亚甲基或亚烷基可以是取代的或非取代的。 In this application, the term "alkylene" generally refers to a saturated straight or branched chain aliphatic hydrocarbon group having 2 hydrogen atoms derived from the same or two different carbon atoms of the parent alkane by removing two hydrogen atoms. residue, which may be a straight or branched chain group containing 1 to 20 carbon atoms, for example, the term "methylene" may refer to a residue derived from a group of 1 carbon atom by removing two hydrogen atoms base. A methylene group may be substituted or unsubstituted, substituted or non-substituted; eg, an alkylene group containing from 1 to 12 carbon atoms, eg, an alkylene group containing from 1 to 6 carbon atoms. Non-limiting examples of alkylene groups include, but are not limited to, methylene ( -CH2- ), 1,1-ethylene (-CH( CH3 )-), 1,2-ethylene ( -CH2) CH 2 )-, 1,1-propylene (-CH(CH 2 CH 3 )-), 1,2-propylene (-CH 2 CH(CH 3 )-), 1,3-propylene ( -CH2CH2CH2- ), 1,4 - Butylene ( -CH2CH2CH2CH2- ) and 1,5 - Butylene ( -CH2CH2CH2CH2CH2- ) Wait. Alkylene may be substituted or unsubstituted, substituted or non-substituted, for example, when substituted, substituents may be substituted at any available point of attachment, which may be independently optionally selected from alkanes group, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy substituted by one or more substituents in the group, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo, such as hydrogen, protium, deuterium, tritium, halogen, -NO 2 , -CN, -OH, -SH, -NH 2 , -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH 2 C(O)H, -S ( O)H, -S(O)2H, -C (O) NH2 , -SO2NH2 , -OC ( O)H, -N(H) SO2H or C1-6 aliphatic . Methylene or alkylene groups can be substituted or unsubstituted.
在本申请中，术语“烯基”通常是指含有一个或多个双键的直链或支链烃基。烯基的示例性实例包括烯丙基、高烯丙基、乙烯基、巴豆基、丁烯基、戊烯基和己烯基等。具有一个以上双键的C 2-6链烯基的示例性实例包括丁二烯基、戊二烯基、己二烯基和己三烯基以及它们的支化形式。不饱和键(双键)的位置可以是在碳链的任何一个位置。烯基可以是取代的或非取代的。 In this application, the term "alkenyl" generally refers to a straight or branched chain hydrocarbon group containing one or more double bonds. Illustrative examples of alkenyl groups include allyl, homoallyl, vinyl, crotyl, butenyl, pentenyl, hexenyl, and the like. Illustrative examples of C2-6 alkenyl groups having more than one double bond include butadienyl, pentadienyl, hexadienyl, and hexatrienyl and branched forms thereof. The position of the unsaturated bond (double bond) can be anywhere in the carbon chain. Alkenyl groups can be substituted or unsubstituted.
在本申请中，术语“亚烯基”通常是指具有从烯烃的碳原子上除去两个氢原子所衍生的残基。例如，可以是亚烯丙基、亚乙烯基、亚丁烯基、亚戊烯基和亚己烯基等。亚烯基可以是取代的或非取代的。In this application, the term "alkenylene" generally refers to residues having two hydrogen atoms removed from the carbon atoms of an olefin. For example, allylene, vinylene, butenylene, pentenylene, hexenylene, and the like can be used. Alkenylene groups can be substituted or unsubstituted.
在本申请中，术语“炔基”通常是指不饱和直链或支链炔基，例如乙炔基、1-丙炔基、炔丙基、丁炔基等。炔基可以是取代的或非取代的。In this application, the term "alkynyl" generally refers to unsaturated straight or branched chain alkynyl groups such as ethynyl, 1-propynyl, propargyl, butynyl, and the like. Alkynyl groups can be substituted or unsubstituted.
在本申请中，术语“亚炔基”通常是指具有从炔烃的碳原子上除去两个氢原子所衍生的残基。例如，可以是亚乙炔基、亚丙炔基、亚炔丙基、亚丁炔基等。亚炔基可以是取代的或非取代的。In this application, the term "alkynylene" generally refers to residues having two hydrogen atoms removed from the carbon atoms of an alkyne. For example, it can be ethynylene, propynylene, propargylene, butynylene and the like. Alkynylene groups can be substituted or unsubstituted.
在本申请中，术语“芳基”通常是指具有芳环上除去一个氢原子所衍生的残基。术语“芳环”可以指具有共轭的π电子体系的6至14元全碳单环或稠合多环(也就是共享毗邻碳原子对的环)，可以为6至10元，例如苯和萘。所述芳环可以稠合于杂芳基、杂环基或环烷基环上，其中与母体结构连接在一起的环为芳基环。芳基可以是取代的或非取代的，当被取代时，取代基可以为一个或多个以下基团，其独立地选自以下组：烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、和杂环烷硫基。芳基可以是取代的或非取代的。In this application, the term "aryl" generally refers to residues having an aromatic ring derived by removing one hydrogen atom. The term "aromatic ring" may refer to a 6- to 14-membered all-carbon monocyclic or fused polycyclic ring (ie, rings that share adjacent pairs of carbon atoms) having a conjugated pi-electron system, and may be 6 to 10 membered, such as benzene and Naphthalene. The aromatic ring can be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring linked to the parent structure is an aryl ring. Aryl may be substituted or unsubstituted, and when substituted, the substituent may be one or more of the following groups independently selected from the group consisting of: alkyl, alkenyl, alkynyl, alkoxy, alkane Thio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio , and heterocycloalkylthio. Aryl groups can be substituted or unsubstituted.
在本申请中，术语“亚芳基”通常是指具有从芳环的碳原子上除去两个氢原子所衍生的残基。例如，可以是亚苯基和亚萘基。亚芳基可以是取代的或非取代的。In this application, the term "arylene" generally refers to a residue having two hydrogen atoms removed from a carbon atom of an aromatic ring. For example, phenylene and naphthylene may be mentioned. Arylene groups can be substituted or unsubstituted.
在本申请中，术语“杂芳基”通常是指具有从杂芳环的碳原子上除去一个氢原子所衍生的残基。术语“杂芳环”指包含1至4个杂原子、5至14个环原子的杂芳族体系，其中杂原子可以选自以下组：氧、硫和氮。杂芳基可以为5至10元，可以为5元或6元，例如呋喃基、噻吩基、吡啶基、吡咯基、N-烷基吡咯基、嘧啶基、吡嗪基、咪唑基、四唑基等。所述杂芳基环可以稠合于芳基、杂环基或环烷基环上，其中与母体结构连接在一起的环为杂芳基环。杂芳基可以是任选取代的或非取代的，当被取代时，取代基可以为一个或多个以下基团，其独立地选自以下组：烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、和杂环 烷硫基。杂芳基可以是取代的或非取代的。In this application, the term "heteroaryl" generally refers to a residue having a hydrogen atom removed from a carbon atom of a heteroaromatic ring. The term "heteroaromatic ring" refers to a heteroaromatic system comprising 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms may be selected from the group consisting of oxygen, sulfur and nitrogen. Heteroaryl can be 5 to 10 membered, 5 membered or 6 membered, such as furanyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazole Base et al. The heteroaryl ring can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is a heteroaryl ring. Heteroaryl groups can be optionally substituted or unsubstituted, and when substituted, the substituents can be one or more of the following groups independently selected from the group consisting of: alkyl, alkenyl, alkynyl, alkoxy group, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, ring Alkylthio, and heterocycloalkylthio. Heteroaryl groups can be substituted or unsubstituted.
在本申请中，术语“亚杂芳基”通常是指具有从杂芳环的碳原子上除去两个氢原子所衍生的残基。例如，可以是亚呋喃基、亚噻吩基、亚吡啶基、亚吡咯基、亚嘧啶基、亚吡嗪基、亚咪唑基、亚四唑基等。亚杂芳基可以是取代的或非取代的。In this application, the term "heteroarylene" generally refers to a residue having two hydrogen atoms removed from a carbon atom of a heteroaromatic ring. For example, it may be a furanylene group, a thienylene group, a pyridylene group, a pyrrolidine group, a pyrimidinylene group, a pyrazinylene group, an imidazolylylene group, a tetrazolium group, and the like. A heteroarylene group can be substituted or unsubstituted.
在本申请中，术语“脂环基”通常是指具有从脂肪环的相同碳原子或多个不同的碳原子上除去氢原子所衍生的残基。术语“环烷”通常指饱和或部分不饱和单环或多环环状烃，碳环包含3至20个碳原子，可以包含3至12个碳原子，可以包含3至10个碳原子，可以包含3至8个碳原子。脂环基的非限制性实例包括环丙烷基、环丁烷基、环戊烷基、环戊烯基、环己烷基、环己烯基、环己二烯基、环庚烷基、环庚三烯基、环辛烷基等；多环碳环可以包括螺环、稠环和桥环的碳环。脂环基可以是取代的或非取代的。在本申请中，术语“碳环基”通常是指具有碳环的碳原子上除去一个氢原子所衍生的残基。术语“碳环”通常指饱和或部分不饱和单环或多环环状烃，碳环包含3至20个碳原子，可以包含3至12个碳原子，可以包含3至10个碳原子，可以包含3至8个碳原子。单环碳环的非限制性实例包括环丙烷、环丁烷、环戊烷、环戊烯、环己烷、环己烯、环己二烯、环庚烷、环庚三烯、环辛烷等；多环碳环可以包括螺环、稠环和桥环的碳环。碳环基可以是取代的或非取代的。在某些情形下，脂环和碳环可以相互替代使用。In this application, the term "alicyclic group" generally refers to a residue having a hydrogen atom removed from the same carbon atom or a plurality of different carbon atoms of an alicyclic ring. The term "cycloalkane" generally refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon, the carbocyclic ring containing 3 to 20 carbon atoms, may contain 3 to 12 carbon atoms, may contain 3 to 10 carbon atoms, may Contains 3 to 8 carbon atoms. Non-limiting examples of alicyclic groups include cyclopropanyl, cyclobutanyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclopentyl Heptatrienyl, cyclooctyl, etc.; polycyclic carbocycles may include spiro, fused, and bridged carbocycles. Alicyclic groups can be substituted or unsubstituted. In this application, the term "carbocyclyl" generally refers to a residue derived from a carbon atom having a carbocyclic ring by removing one hydrogen atom. The term "carbocycle" generally refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon, the carbocycle contains 3 to 20 carbon atoms, may contain 3 to 12 carbon atoms, may contain 3 to 10 carbon atoms, may Contains 3 to 8 carbon atoms. Non-limiting examples of monocyclic carbocycles include cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexene, cyclohexadiene, cycloheptane, cycloheptatriene, cyclooctane etc.; polycyclic carbocycles may include spiro, fused and bridged carbocycles. Carbocyclyl groups can be substituted or unsubstituted. Alicyclic and carbocyclic may be used interchangeably in some cases.
在本申请中，术语“部分不饱和的”通常是指环状结构中环分子间至少含一个双键或三键。术语“部分不饱和”涵盖带有多处不饱和的环状结构，但并非意在包括本申请所定义的芳环或杂芳环。术语"不饱和的"表示部分具有一个或多个不饱和度。In this application, the term "partially unsaturated" generally refers to a cyclic structure containing at least one double or triple bond between the ring molecules. The term "partially unsaturated" encompasses cyclic structures with multiple unsaturations, but is not intended to include aromatic or heteroaromatic rings as defined herein. The term "unsaturated" means that the moiety has one or more degrees of unsaturation.
在本申请中，术语“亚脂环基”通常是指具有从脂环的碳原子上除去两个氢原子所衍生的残基。例如，可以是亚环丙烷基、亚环丁烷基、亚环戊烷基、亚环戊烯基、亚环己烷基、亚环己烯基、亚环己二烯基、亚环庚烷基、亚环庚三烯基、亚环辛烷基等；多环碳环可以包括螺环、稠环和桥环的碳环。亚脂环基可以是取代的或非取代的。In this application, the term "alicyclic group" generally refers to a residue having two hydrogen atoms removed from a carbon atom of an alicyclic ring. For example, it can be cyclopropene, cyclobutane, cyclopentylene, cyclopentenylene, cyclohexylene, cyclohexenylene, cyclohexadienylene, cycloheptane cycloheptatrienyl, cyclooctylene, etc.; polycyclic carbocycles may include spiro, fused, and bridged carbocycles. Alicyclic groups can be substituted or unsubstituted.
在本申请中，术语“脂杂环基”通常是指稳定的不具有芳香性的3元-7元单环碳环结构，融合的7元-10元双环杂环结构或桥联的6元-10元双环杂环结构，这些环状结构即可以是饱和的，也可以是部分饱和的，除碳原子外，这些环状结构中还含有一个或多个杂原子，其中杂原子可以选自以下组：氧、硫和氮。例如是含有1-4个上述定义的杂原子。当用来表示脂杂环环状结构上的原子时，术语“氮”可以包括发生过取代反应的氮。例如，脂杂环基可以包含“杂环烷基”，杂环烷基可以指稳定的不具有芳香性的3元-7元单环烷结构，融合的7元-10元双环杂环结构或桥联的6元-10元双环杂环结构，除碳原子外，这些环状结构中还含有一 个或多个杂原子，其中杂原子可以选自以下组：氧、硫和氮。例如是含有1-4个上述定义的杂原子。杂环烷基可以是取代的或非取代的。脂杂环基可以是取代的或非取代的。In this application, the term "alicyclic heterocyclyl" generally refers to stable non-aromatic 3- to 7-membered monocyclic carbocyclic structures, fused 7- to 10-membered bicyclic heterocyclic structures or bridged 6-membered -10-membered bicyclic heterocyclic structures, these cyclic structures can be either saturated or partially saturated, in addition to carbon atoms, these cyclic structures also contain one or more heteroatoms, wherein the heteroatoms can be selected from The following groups: oxygen, sulfur and nitrogen. For example, it contains 1-4 heteroatoms as defined above. The term "nitrogen" when used to refer to an atom on an alicyclic ring structure may include nitrogen that has undergone a substitution reaction. For example, a heteroalicyclic group may include "heterocycloalkyl", which may refer to a stable non-aromatic 3- to 7-membered monocycloalkane structure, a fused 7- to 10-membered bicyclic heterocyclic structure or Bridged 6- to 10-membered bicyclic heterocyclic structures containing, in addition to carbon atoms, one or more heteroatoms, wherein the heteroatoms may be selected from the group consisting of oxygen, sulfur and nitrogen. For example, it contains 1-4 heteroatoms as defined above. Heterocycloalkyl can be substituted or unsubstituted. Alicyclic groups may be substituted or unsubstituted.
在本申请中，术语“亚脂杂环基”通常是指具有从脂杂环的碳原子上除去两个氢原子所衍生的残基。亚脂杂环基可以是取代的或非取代的。In this application, the term "alicyclic heterocyclyl" generally refers to a residue having two hydrogen atoms removed from a carbon atom of an alicyclic ring. Aliphatic heterocyclyl groups can be substituted or unsubstituted.
在本申请中，术语“任选”或“任选地”通常是指随后所描述的事件或环境可以但不必发生，该说明包括该事件或环境发生或不发生地场合。例如，“任选被烷基取代的杂环基团”意味着烷基可以但不必须存在，该说明可以包括杂环基团被烷基取代的情形和杂环基团不被烷基取代的情形。In this application, the terms "optional" or "optionally" generally mean that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs or does not occur. For example, "a heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may, but need not, be present, and the specification may include both instances where the heterocyclic group is substituted with an alkyl group and where the heterocyclic group is not substituted with an alkyl group. situation.
在本申请中，术语“取代的”通常是指基团中的一个或多个氢原子，例如为最多5个，例如为1～3个氢原子彼此独立地被相应数目的取代基取代。取代基仅处在它们的可能的化学位置，本领域技术人员能够在不付出过多努力的情况下确定(通过实验或理论)可能或不可能的取代。例如，具有游离氢的氨基或羟基与具有不饱和(如烯属)键的碳原子结合时可能是不稳定的。In the present application, the term "substituted" generally means that one or more hydrogen atoms in a group, eg up to 5, eg 1 to 3 hydrogen atoms, independently of one another, are substituted by the corresponding number of substituents. Substituents are only in their possible chemical positions, and those skilled in the art can determine (either experimentally or theoretically) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups with free hydrogens may be unstable when combined with carbon atoms with unsaturated (eg, olefinic) bonds.
在本申请中，如本领域技术人员可知的，“烷基”、“烯基”、“环烷基”等之类的术语可以在名称前加一个标识表示在特定情况下基团中存在的原子数，例如，C 1-C 4烷基，C 3-C 7环烷氧基，C 1-C 4烷基羰基氨基等，“C”后所跟下标数字表示在基团中存在的碳原子数。例如，C 3烷基是指具有三个碳原子的烷基(例如，正丙基，异丙基)；C 1-10中，基团的成员可具有落入1-10范围内的任何数目的碳原子。 In this application, terms such as "alkyl,""alkenyl,""cycloalkyl," etc. may be preceded by a designation to indicate the presence of The number of atoms, for example, C 1 -C 4 alkyl, C 3 -C 7 cycloalkoxy, C 1 -C 4 alkylcarbonylamino, etc., "C" followed by a subscript number indicates the number of atoms present in the group number of carbon atoms. For example, C3 alkyl refers to an alkyl group having three carbon atoms (eg, n-propyl, isopropyl); in C1-10 , the members of the group can have any number falling within the range of 1-10 of carbon atoms.
基团中的一个或多个氢原子，例如为最多5个，例如为1～3个氢原子彼此独立地被相应数目的取代基取代。取代基仅处在它们的可能的化学位置，本领域技术人员能够在不付出过多努力的情况下确定(通过实验或理论)可能或不可能的取代。例如，具有游离氢的氨基或羟基与具有不饱和(如烯属)键的碳原子结合时可能是不稳定的。One or more hydrogen atoms in the group, for example up to 5, for example 1 to 3 hydrogen atoms, independently of one another, are substituted by the corresponding number of substituents. Substituents are only in their possible chemical positions, and those skilled in the art can determine (either experimentally or theoretically) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups with free hydrogens may be unstable when combined with carbon atoms with unsaturated (eg, olefinic) bonds.
在本申请中，术语“化合物”通常指具有两种或两种以上不同元素的物质。例如，本申请的化合物可以是有机化合物，例如本申请的化合物可以是分子量500以下的化合物，可以是分子量1000以下的化合物，也可以是分子量1000以上的化合物，也可以是10000以上、100000以上的化合物。在本申请中，化合物还可以是指通过化学键相连的化合物，例如可以是一个或多个分子量1000以下的分子通过化学键与生物大分子相连的化合物，所述生物大分子可以是高聚糖、蛋白、核酸、多肽等。例如本申请的化合物可以包括蛋白质与一个或多个分子量1000以下的分子相连的化合物，可以是包括蛋白质与一个或多个分子量10000以下的分子相连的化合物，可以是包括蛋白质与一个或多个分子量100000以下的分子相连的 化合物。In this application, the term "compound" generally refers to a substance having two or more different elements. For example, the compound of the present application may be an organic compound. For example, the compound of the present application may be a compound with a molecular weight of 500 or less, a compound with a molecular weight of 1,000 or less, or a compound with a molecular weight of 1,000 or more, or a compound of 10,000 or more and 100,000 or more. compound. In this application, a compound may also refer to a compound connected by chemical bonds, for example, it may be a compound in which one or more molecules with a molecular weight below 1000 are connected with a biological macromolecule by chemical bonds, and the biological macromolecule may be a high polysaccharide, a protein , nucleic acids, peptides, etc. For example, the compounds of the present application may include compounds in which proteins are linked to one or more molecules with a molecular weight of less than 1000; A compound with less than 100,000 molecules linked together.
在本申请中，术语“包含”通常是指包括明确指定的特征，但不排除其他要素。术语“以上”、“以下”通常是指包含本数的情况。In this application, the term "comprising" generally means including the expressly specified features, but not excluding other elements. The terms "above" and "below" generally refer to the inclusion of this number.
在本申请中，术语“约”通常是指在指定数值以上或以下0.5％-10％的范围内变动，例如在指定数值以上或以下0.5％、1％、1.5％、2％、2.5％、3％、3.5％、4％、4.5％、5％、5.5％、6％、6.5％、7％、7.5％、8％、8.5％、9％、9.5％、或10％的范围内变动。In this application, the term "about" generally refers to a range of 0.5%-10% above or below the specified value, such as 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10%.
在本申请中，本申请的化合物包含化合物的其互变异构体、内消旋体、外消旋体、对映异构体、和/或非对映异构体。在本申请中，术语“非对映异构体”通常是指具有两个或更多个手性中心并且其分子不是彼此的镜像的立体异构体。非对映异构体可以具有不同的物理性质，例如、熔点、沸点、波谱性质和反应性。在本申请中，术语“互变异构体”或“互变异构形式”可互换使用，通常是指可通过低能垒(low energy barrier)互相转化的不同能量的结构异构体。例如，质子互变异构体(protontautomer)(也称为质子移变互变异构体(prototropic tautomer))包括通过质子迁移进行的互相转化，诸如酮-烯醇异构化和亚胺-烯胺异构化。价键互变异构体(valence tautomer)包括通过一些成键电子的重组进行的互相转化。在本申请中，术语“内消旋体”通常是指分子内含有不对称性的原子，但具有对称因素而使分子内总旋光度为零。术语"外消旋体"或"外消旋混合物"是指由等摩尔量的两种对映异构体物质构成的组合物。In the present application, the compounds of the present application include tautomers, mesomers, racemates, enantiomers, and/or diastereomers of the compounds. In this application, the term "diastereomer" generally refers to stereoisomers that have two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers can have different physical properties, eg, melting points, boiling points, spectral properties, and reactivities. In this application, the terms "tautomer" or "tautomeric form" are used interchangeably and generally refer to structural isomers of different energies that can be interconverted through a low energy barrier. For example, protontautomers (also known as prototropic tautomers) include interconversions by migration of protons, such as keto-enol isomerization and imine-ene Amine isomerization. Valence tautomers include interconversions by recombination of some of the bonding electrons. In this application, the term "mesome" generally refers to atoms that contain asymmetry in the molecule, but have a symmetry factor such that the total optical rotation in the molecule is zero. The term "racemate" or "racemic mixture" refers to a composition consisting of two enantiomeric species in equimolar amounts.
在本申请中，本申请的化合物的某些原子可能以一种以上的同位素形式出现。例如，氢可能以氕( 1H)、氘( 2H)和氚( 3H)的形式存在，碳可能以三种不同的同位素( 12C、 13C和 14C)自然存在。可并入本申请化合物中的同位素示例还包括但不限于 15N、 18O、 17O、 18F、 32P、 33P、 129I、 131I、 123I、 124I、 125I，或者类似的同位素。因此，相对于这些同位素的自然丰度，本申请的化合物可富集在一种或多种这些同位素中。如本领域技术人员所知，此类同位素富集化合物可用于多种用途。例如，用重同位素如氘( 2H)替代可能会提供某些治疗优势，这可以是由于更高的代谢稳定性。例如，氘( 2H)的自然丰度约为0.015％。因此，自然界中大约每6500个氢原子，就有一个氘原子。因此，本申请的含氘化合物在一个或多个位置(视情况而定)的氘丰度大于0.015％。除非另有指明，否则本申请所述的结构还可以包括仅在是否存在一个或多个同位素富集原子方面存在差别的化合物。举例而言，除了氢原子被氘或氚所取代，或碳原子被碳13或碳14所取代之外，其余部分均与本申请结构一致的化合物均在本申请的范围之内。 In the present application, certain atoms of the compounds of the present application may occur in more than one isotopic form. For example, hydrogen may exist as protium ( 1 H), deuterium ( 2 H), and tritium ( 3 H), and carbon may exist naturally in three different isotopes ( 12 C, 13 C, and 14 C). Examples of isotopes that can be incorporated into the compounds of the present application also include, but are not limited to, 15 N, 18 O, 17 O, 18 F, 32 P, 33 P, 129 I, 131 I, 123 I, 124 I, 125 I, or the like of isotopes. Thus, the compounds of the present application may be enriched in one or more of these isotopes relative to their natural abundance. Such isotopically enriched compounds can be used for a variety of purposes, as known to those skilled in the art. For example, substitution with heavy isotopes such as deuterium ( 2 H) may offer certain therapeutic advantages, which may be due to greater metabolic stability. For example, the natural abundance of deuterium ( 2 H) is about 0.015%. Therefore, for every 6,500 hydrogen atoms in nature, there is one deuterium atom. Accordingly, the deuterium-containing compounds of the present application have a deuterium abundance greater than 0.015% at one or more positions (as the case may be). Unless otherwise indicated, structures described herein may also include compounds that differ only in the presence or absence of one or more isotopically enriched atoms. For example, except that the hydrogen atom is replaced by deuterium or tritium, or the carbon atom is replaced by carbon 13 or carbon 14, the compounds whose structure is consistent with the present application are all within the scope of the present application.
在本申请中，术语“药物组合物”通常是指含有一种或多种本申请所述化合物或其生理学 上/可药用的盐或前体药物与其他化学组分的混合物，以及其他组分例如生理学/可药用的载体和赋形剂。药物组合物可以是促进对生物体的给药，利于活性成分的吸收进而发挥生物活性。常规的药物组合物的制备可以见本领域常用技术。In this application, the term "pharmaceutical composition" generally refers to a mixture containing one or more of the compounds described herein, or a physiologically/pharmaceutically acceptable salt or prodrug thereof, and other chemical components, as well as other Such as physiological/pharmaceutically acceptable carriers and excipients. The pharmaceutical composition can facilitate the administration to the organism, facilitate the absorption of the active ingredient and then exert the biological activity. The preparation of conventional pharmaceutical compositions can be found in the techniques commonly used in the art.
在本申请中，术语“药学上可接受的盐”或“可药用的盐”通常是指本申请化合物或配体-药物偶联物的盐，或本申请中所述的化合物的盐，这类盐用于哺乳动物体内时可以具有安全性和/或有效性，且可以具有应有的生物活性，本申请抗体-抗体药物偶联化合物可以与酸形成盐，药学上可接受的盐的非限制性实例包括：盐酸盐、氢溴酸盐、氢碘酸盐、硫酸盐、硫酸氢盐、柠檬酸盐、乙酸盐、琥珀酸盐、抗坏血酸盐、草酸盐、硝酸盐、梨酸盐、磷酸氢盐、磷酸二氢盐、水杨酸盐、柠檬酸氢盐、酒石酸盐、马来酸盐、富马酸盐、甲酸盐、苯甲酸盐、甲磺酸盐、乙磺酸盐、苯磺酸盐、或对甲苯磺酸盐。In this application, the term "pharmaceutically acceptable salt" or "pharmaceutically acceptable salt" generally refers to a salt of a compound or ligand-drug conjugate of the present application, or a salt of a compound described in the present application, Such salts can be safe and/or effective when used in mammals, and can have due biological activity, and the antibody-antibody drug conjugate compounds of the present application can form salts with acids, and the pharmaceutically acceptable salts are salts. Non-limiting examples include: hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, citrate, acetate, succinate, ascorbate, oxalate, nitrate, pear Acid, hydrogen phosphate, dihydrogen phosphate, salicylate, hydrogen citrate, tartrate, maleate, fumarate, formate, benzoate, mesylate, ethyl acetate Sulfonate, benzenesulfonate, or p-toluenesulfonate.
在本申请中，术语“偶联物”通常是指本申请的化合物通过一个或多个化学反应而制备的化合物，或者通过诸如桥接(bridge)，间隔物(spacer)，或连接部分等的一个或多个连接结构彼此连接。In the present application, the term "conjugate" generally refers to a compound of the present application prepared by one or more chemical reactions, or by one such as a bridge, spacer, or linking moiety, etc. or multiple connecting structures are connected to each other.
在本申请中，术语“药学上可接受的载体”通常是指给予治疗剂，例如抗体或多肽、基因和其它治疗剂的载体。该术语指本身不诱导对接受组合物的个体有害的抗体产生并且可以给予而不产生过度毒性的任何药物载体。例如，药学上可接受的载体(carrier)可以与基因工程中用于包含目标基因的核酸载体(vector)相区分。合适的载体可以是大的、代谢缓慢的大分子，例如蛋白质、多糖、聚乳酸、聚乙醇酸、多聚氨基酸、氨基酸共聚物、脂质聚集物和灭活的病毒颗粒。本领域技术人员熟知这些载体。治疗组合物中药学上可接受的载体可包括液体，例如水、盐水、甘油和乙醇。这些载体中也可存在辅助物质，例如润湿剂或乳化剂、pH缓冲物质等。In this application, the term "pharmaceutically acceptable carrier" generally refers to a carrier for administration of therapeutic agents, such as antibodies or polypeptides, genes and other therapeutic agents. The term refers to any pharmaceutical carrier that itself does not induce the production of antibodies detrimental to the individual receiving the composition and can be administered without undue toxicity. For example, a pharmaceutically acceptable carrier can be distinguished from a nucleic acid vector used in genetic engineering to contain a gene of interest. Suitable carriers can be large, slowly metabolized macromolecules such as proteins, polysaccharides, polylactic acid, polyglycolic acid, polyamino acids, amino acid copolymers, lipid aggregates, and inactivated viral particles. Those skilled in the art are familiar with these vectors. Pharmaceutically acceptable carriers in therapeutic compositions can include liquids such as water, saline, glycerol and ethanol. Auxiliary substances such as wetting or emulsifying agents, pH buffering substances and the like can also be present in these carriers.
在本申请中，术语“Trop2”、“TROP2”通常是指单程跨膜I型细胞膜蛋白。在本申请中，术语“Trop2”还可涵盖Trop 2的同源物、变体和同工型，包括剪接同工型。术语“Trop”还包括具有Trop 2同源物、变体和同工型中的一个或多个序列的蛋白，以及该序列的片段，只要是该变体蛋白(包括同工型)。Trop2可以是人Trop2。例如，Uniprot登录号P09758提供了Trop2和序列的描述。In this application, the terms "Trop2", "TROP2" generally refer to a single-pass transmembrane type I cell membrane protein. In this application, the term "Trop2" may also encompass homologues, variants and isoforms of Trop 2, including splice isoforms. The term "Trop" also includes proteins having one or more sequences of Trop 2 homologues, variants and isoforms, as well as fragments of such sequences, so long as the variant protein (including isoforms). Trop2 can be human Trop2. For example, Uniprot Accession No. P09758 provides a description of Trop2 and sequences.
在本申请中，术语“HER2”通常是指人表皮生长因子受体2(HER2)。例如，术语“HER2”指来自任何人来源的任何天然HER2。该术语还涵盖“全长”和未加工的HER2以及源自细胞中加工的任何形式的HER2(例如成熟蛋白)。该术语还涵盖HER2的天然发生变体和同等型，例如剪接变体或等位变体。例如，Uniprot登录号P04626提供了HER2和序列的描述。In this application, the term "HER2" generally refers to human epidermal growth factor receptor 2 (HER2). For example, the term "HER2" refers to any native HER2 from any human source. The term also encompasses "full-length" and unprocessed HER2 as well as any form of HER2 derived from processing in a cell (eg, the mature protein). The term also encompasses naturally occurring variants and isoforms of HER2, such as splice variants or allelic variants. For example, Uniprot Accession No. P04626 provides a description of HER2 and sequences.
在本申请中，术语“Nectin-4”通常是指粘附分子4。例如，术语“Nectin-4”指来自任何人来源的任何天然Nectin-4。该术语还涵盖“全长”和未加工的Nectin-4以及源自细胞中加工的任何形式的Nectin-4(例如成熟蛋白)。该术语还涵盖Nectin-4的天然发生变体和同等型，例如剪接变体或等位变体。例如，Uniprot登录号Q96NY8提供了Nectin-4和序列的描述。In this application, the term "Nectin-4" generally refers to adhesion molecule 4. For example, the term "Nectin-4" refers to any native Nectin-4 from any human source. The term also encompasses "full-length" and unprocessed Nectin-4 as well as any form of Nectin-4 derived from processing in a cell (eg, mature protein). The term also encompasses naturally occurring variants and isoforms of Nectin-4, such as splice variants or allelic variants. For example, Uniprot Accession No. Q96NY8 provides a description of Nectin-4 and sequences.
在本申请中，术语“嵌合抗体(chimeric antibody)”通常是指鼠源性抗体的可变区与人抗体的恒定区融合而成的抗体，可以减轻鼠源性抗体诱发的免疫应答反应。建立嵌合抗体，可以建立分泌鼠源性特异性单抗的杂交瘤，然后从鼠杂交瘤细胞中克隆可变区基因，可以根据需要克隆人抗体的恒定区基因，将鼠可变区基因与人恒定区基因连接成嵌合基因后插入表达载体中，可以在真核系统或原核系统中表达嵌合抗体分子。In this application, the term "chimeric antibody" generally refers to an antibody in which the variable region of a murine antibody is fused with the constant region of a human antibody, which can alleviate the immune response induced by the murine antibody. To establish a chimeric antibody, a hybridoma that secretes a mouse-specific monoclonal antibody can be established, and then the variable region gene can be cloned from the mouse hybridoma cell, and the constant region gene of the human antibody can be cloned according to the needs. The human constant region gene is connected into a chimeric gene and inserted into an expression vector, and the chimeric antibody molecule can be expressed in a eukaryotic system or a prokaryotic system.
在本申请中，术语“人源化抗体(humanized antibody)”，也称为CDR移植抗体(CDR-grafted antibody)，通常是指将鼠的CDR序列移植到人的抗体可变区框架，即不同类型的人种系抗体框架序列中产生的抗体。可以克服嵌合抗体由于携带大量鼠蛋白成分，从而诱导的异源性反应。此类构架序列可以从包括种系抗体基因序列的公共DNA数据库或公开的参考文献获得。如人重链和轻链可变区基因的种系DNA序列可以在“VBase”人种系序列数据库。In this application, the term "humanized antibody", also known as CDR-grafted antibody, generally refers to the grafting of murine CDR sequences into the framework of human antibody variable regions, i.e. different Types of human germline antibody framework sequences produced in antibodies. The heterologous reaction induced by chimeric antibodies can be overcome because they carry a large amount of murine protein components. Such framework sequences can be obtained from public DNA databases or published references that include germline antibody gene sequences. For example, the germline DNA sequences of the human heavy and light chain variable region genes can be found in the "VBase" human germline sequence database.
在本申请中，术语“全人源抗体”、“全人抗体”或“完全人源抗体”，也称“全人源单克隆抗体”，其抗体的可变区和恒定区可以都是人源的，去除免疫原性和毒副作用。单克隆抗体的发展经历了四个阶段，分别为：鼠源性单克隆抗体、嵌合性单克隆抗体、人源化单克隆抗体和全人源单克隆抗体。本申请所述抗体或配体可以为全人源单克隆抗体。全人抗体制备的相关技术可以为：人杂交瘤技术、EBV转化B淋巴细胞技术、噬菌体显示技术(phage display)、转基因小鼠抗体制备技术(transgenic mouse)和单个B细胞抗体制备技术等。In this application, the term "fully human antibody", "fully human antibody" or "fully human antibody", also referred to as "fully human monoclonal antibody", the variable region and constant region of the antibody can be both human source, to remove immunogenicity and toxic side effects. The development of monoclonal antibodies has gone through four stages, namely: murine monoclonal antibodies, chimeric monoclonal antibodies, humanized monoclonal antibodies and fully human monoclonal antibodies. The antibodies or ligands described herein may be fully human monoclonal antibodies. Related technologies for the preparation of fully human antibodies include: human hybridoma technology, EBV transformation of B lymphocytes, phage display technology, transgenic mouse antibody preparation technology, and single B cell antibody preparation technology.
在本申请中，术语“CDR”通常是指抗体的可变结构域内主要促成抗原结合的6个高变区之一。所述6个CDR的最常用的定义之一由Kabat E.A.等人，Chothia等人和MacCallum等人提供。如本申请中使用的，CDR的Kabat定义可以应用于轻链可变结构域的CDR1、CDR2和CDR3(CDR L1、CDR L2、CDR L3或L1、L2、L3)，以及重链可变结构域的CDR1、CDR2和CDR3(CDR H1、CDR H2、CDR H3或H1、H2、H3)。In this application, the term "CDR" generally refers to one of the six hypervariable regions within the variable domain of an antibody that primarily contribute to antigen binding. One of the most commonly used definitions of the 6 CDRs is provided by Kabat E.A. et al., Chothia et al. and MacCallum et al. As used in this application, the Kabat definition of CDRs can be applied to CDR1, CDR2 and CDR3 (CDR L1, CDR L2, CDR L3 or L1, L2, L3) of the light chain variable domains, as well as the heavy chain variable domains of CDR1, CDR2 and CDR3 (CDR H1, CDR H2, CDR H3 or H1, H2, H3).
在本申请中，术语“能够与巯基偶联的基团”通常是指所述化合物A具有巯基，所述化合物B具有能够与巯基偶联的基团，化合物B通过能够与巯基偶联的基团与化合物A的巯基反应，可以以此实现化合物A与化合物B的连接。In the present application, the term "group capable of coupling with a thiol group" generally means that the compound A has a thiol group, the compound B has a group capable of coupling with a thiol group, and the compound B has a group capable of coupling with a thiol group through a group capable of coupling with a thiol group. The group reacts with the sulfhydryl group of compound A, which can realize the connection between compound A and compound B.
在本申请中，术语“连接体”通常是指一端与一个基团连接而另一端与另一个基团相连的化学结构片段或键，也可以连接其他连接体后再与药物和/或配体相连。所述直接或间接连 接配体可以是指所述基团通过共价键直接连接配体，也可以是通过连接体连接配体。例如，连接体可以是本申请所述的Q 1所示的结构。例如，可以使用包含酸不稳定接头结构(例如腙)、蛋白酶敏感(例如肽酶敏感)接头结构、光不稳定接头结构、二甲基接头结构、或含二硫化物接头结构的化学结构片段或键作为连接体。 In this application, the term "linker" usually refers to a chemical structural fragment or bond with one end connected to one group and the other end connected to another group. It can also be connected to other linkers and then to drugs and/or ligands. connected. The direct or indirect linking of the ligand may refer to that the group is directly linked to the ligand through a covalent bond, or may be linked to the ligand through a linker. For example, the linker can be the structure shown by Q 1 described in this application. For example, chemical fragments comprising acid-labile linker structures (eg, hydrazones), protease-sensitive (eg, peptidase-sensitive) linker structures, photolabile linker structures, dimethyl linker structures, or disulfide-containing linker structures may be used or bond as a linker.
在本申请中，术语“连接基团”通常是指具有与另一个基团相连的能力的基团。例如，具有连接基团的化合物，可以通过该连接基团与另一个基团的偶联反应，实现该化合物与另一个基团的连接。例如，马来酰亚胺基团可以作为连接基团。In this application, the term "linking group" generally refers to a group that has the ability to attach to another group. For example, for a compound with a linking group, the linking of the compound with another group can be achieved through a coupling reaction between the linking group and another group. For example, a maleimide group can serve as the linking group.
在本申请中，术语“药物单元”通常是指直接或间接缀合抗体或抗原结合片段以形成免疫缀合物的化学部分。例如，“药物单元”包括但不限于文中描述的抗肿瘤活性的化合物。例如药物单元包括拓扑异构酶抑制剂。In this application, the term "drug unit" generally refers to a chemical moiety that directly or indirectly conjugates an antibody or antigen-binding fragment to form an immunoconjugate. For example, a "drug unit" includes, but is not limited to, compounds described herein with anti-tumor activity. For example, the drug unit includes a topoisomerase inhibitor.
在本申请中，术语“抗肿瘤活性的化合物”通常是指具有使肿瘤细胞的增殖速率、存活力或转移活性的降低的能力的化合物。例如，抗肿瘤活性可以由治疗期间出现的异常细胞的生长速率减少或肿瘤尺寸稳定或缩减显示的，或与在无治疗情况下对照相比因治疗所致的存活期更长显示的。抗肿瘤活性可使用公认的体外或体内肿瘤模型评估，例如异种移植模型。In the present application, the term "compound with antitumor activity" generally refers to a compound having the ability to reduce the proliferation rate, viability or metastatic activity of tumor cells. For example, anti-tumor activity may be manifested by a reduction in the growth rate of abnormal cells or by stabilization or shrinkage of tumor size during treatment, or by longer survival due to treatment compared to a control in the absence of treatment. Antitumor activity can be assessed using recognized in vitro or in vivo tumor models, such as xenograft models.
在本发明的部分实施方案中，偶联物中的生物活性分子为具体有抗肿瘤活性的化合物，具体例如：放射性同位素，例如At 211、I 131、I 125、Y 90、Re 186、Re 188、Sm 153、Bi 212、P 32、Pb 212或Lu的放射性同位素；金属配合物，例如金属铂配合物(如奥沙利铂)或金属金配合物；糖肽类抗生素，例如博来霉素或平阳霉素；拓扑异构酶抑制剂；干扰DNA合成的药物，例如甲氨蝶呤、5-氟尿嘧啶、阿糖胞苷、吉西他滨、巯嘌呤、喷司他丁、氟达拉滨、克拉屈滨或奈拉滨等；作用于结构蛋白的药物，例如微管蛋白抑制剂(诸如长春花生物碱类、长春新碱、长春碱、紫杉醇类、美登素类、澳瑞他汀类、Tubulysin B或艾瑞布林等)；肿瘤信号通路抑制剂，例如丝氨酸/苏氨酸激酶抑制剂、酪氨酸激酶抑制剂、天冬氨酸激酶抑制剂或组氨酸激酶抑制剂等；蛋白酶体抑制剂；表观遗传相关靶点抑制剂；肿瘤新生血管生成抑制剂；细胞周期蛋白抑制剂； In some embodiments of the present invention, the biologically active molecule in the conjugate is a compound with specific anti-tumor activity, for example, a radioisotope, such as At 211 , I 131 , I 125 , Y 90 , Re 186 , Re 188 , Sm 153 , Bi 212 , P 32 , Pb 212 , or radioisotopes of Lu; metal complexes, such as metal platinum complexes (such as oxaliplatin) or metal gold complexes; glycopeptide antibiotics, such as bleomycin or pingyangmycin; topoisomerase inhibitors; drugs that interfere with DNA synthesis, such as methotrexate, 5-fluorouracil, cytarabine, gemcitabine, mercaptopurine, pentostatin, fludarabine, cladrix Drugs acting on structural proteins, such as tubulin inhibitors (such as vinca alkaloids, vincristine, vinblastine, paclitaxel, maytansine, auristatin, Tubulysin B or eribulin, etc.); tumor signaling pathway inhibitors, such as serine/threonine kinase inhibitors, tyrosine kinase inhibitors, aspartate kinase inhibitors or histidine kinase inhibitors, etc.; proteasome inhibition Agents; Epigenetic-related Target Inhibitors; Tumor Angiogenesis Inhibitors; Cyclin Inhibitors;
在本申请中，术语“拓扑异构酶抑制剂”通常是指包括拓扑异构酶I抑制剂和拓扑异构酶II抑制剂的化合物或其衍生物。拓扑异构酶I抑制剂的实例包括但不限于喜树碱及其类似物；拓扑异构酶II抑制剂(诸如放线菌素D、阿霉素、多柔比星、多卡米星、柔红霉素、米托蒽醌、鬼臼毒素或依托泊苷等)。拓扑异构酶(topoisomerase)可以是指通过切断DNA的一条或两条链中的磷酸二酯键，然后重新缠绕和封口来更正DNA连环数的酶。In this application, the term "topoisomerase inhibitor" generally refers to compounds or derivatives thereof that include topoisomerase I inhibitors and topoisomerase II inhibitors. Examples of topoisomerase I inhibitors include, but are not limited to, camptothecin and its analogs; topoisomerase II inhibitors (such as actinomycin D, doxorubicin, doxorubicin, docarmycin, daunorubicin, mitoxantrone, podophyllotoxin or etoposide, etc.). A topoisomerase may refer to an enzyme that corrects the number of concatenated chains of DNA by cleaving phosphodiester bonds in one or both strands of DNA, followed by rewinding and sealing.
在本申请中，术语“喜树碱类似物”通常是指与喜树碱结构类似或衍生自喜树碱的化合物。例如，喜树碱的结构可以在CAS登录号7689-03-4中记载。例如，喜树碱类似物可以是指伊沙替康(Exatecan，CAS登录号171335-80-1)或贝洛替康(Belotecan，CAS登录号256411-32-2)。术语“非喜树碱类拓扑异构酶I抑制剂”通常是指吲哚咔唑类、茚并异喹啉酮类、苯并菲啶类以及二苯并萘啶酮类具有拓扑异构酶I抑制活性的杂环分子，主要指Genz-644282(CAS登录号529488-28-6)。In this application, the term "camptothecin analog" generally refers to compounds that are structurally similar to or derived from camptothecin. For example, the structure of camptothecin can be described in CAS Accession No. 7689-03-4. For example, a camptothecin analog may refer to ixatecan (Exatecan, CAS Accession No. 171335-80-1) or Belotecan (Belotecan, CAS Accession No. 256411-32-2). The term "non-camptothecin-type topoisomerase I inhibitors" generally refers to indolecarbazoles, indenoisoquinolinones, triphenanthridines, and dibenzonaphthyridinones with topoisomerases I heterocyclic molecules with inhibitory activity mainly refer to Genz-644282 (CAS accession number 529488-28-6).
在本申请中，术语与某靶点“表达相关”的疾病通常是指该疾病的发生和/或进展与该靶点的表达水平有关联。例如，相对于来自组织或器官的正常细胞的表达水平，来自疾病区如患者的特定组织或器官内的细胞中某个靶点的表达水平提高，即高表达的。或者例如，相对于来自组织或器官的正常细胞的表达水平，来自疾病区如患者的特定组织或器官内的细胞中某个靶点的表达水平降低，即低表达的。或者例如，来自疾病区如患者的特定组织或器官内的细胞表达某个靶点，即阳性的。或者例如，来自疾病区如患者的特定组织或器官内的细胞不表达某个靶点，即阴性的。例如，靶点表达的特征可以由本领域已知的标准测定确定。In the present application, the term "expression-related" disease of a target generally means that the occurrence and/or progression of the disease is associated with the expression level of the target. For example, the expression level of a target in cells from a disease area, such as within a particular tissue or organ of a patient, is increased, ie, highly expressed, relative to the expression level of normal cells from the tissue or organ. Or, for example, the expression level of a certain target in cells from a disease area, such as a particular tissue or organ of a patient, is reduced, ie, underexpressed, relative to the expression level of normal cells from the tissue or organ. Or, for example, cells from a disease area, such as within a particular tissue or organ of a patient, express a certain target, ie, are positive. Or, for example, cells from a disease area, such as a particular tissue or organ of a patient, do not express a certain target, ie, are negative. For example, target expression can be characterized by standard assays known in the art.
在本申请中，术语“有效量”通常是指治疗剂治疗、缓解或预防目标疾病或状况的量，或是表现出可检测的治疗或预防效果的量。对于某一对象的精确有效量取决于该对象的体型和健康状况、病症的性质和程度、以及选择给予的治疗剂和/或治疗剂的组合。因此，预先指定准确的有效量是没用的。然而，对于某给定的状况而言，可以用常规实验来确定该有效量，临床医师是能够判断出来的。In this application, the term "effective amount" generally refers to the amount of a therapeutic agent that treats, ameliorates, or prevents a target disease or condition, or an amount that exhibits a measurable therapeutic or prophylactic effect. The precise effective amount for a subject depends on the size and health of the subject, the nature and extent of the disorder, and the therapeutic agent and/or combination of therapeutic agents selected for administration. Therefore, it is useless to prespecify the exact effective amount. However, for a given situation, routine experimentation can be used to determine the effective amount, as is the judgment of the clinician.
除非特别说明，本申请中，所有出现的化合物均意在包括所有可能的光学异构体，如单一手性的化合物，或各种不同手性化合物的混合物(即外消旋体)。本申请的所有化合物之中，各手性碳原子可以任选地为R构型或S构型，或R构型和S构型的混合物。Unless otherwise specified, in this application, all occurrences of compounds are intended to include all possible optical isomers, such as single chiral compounds, or mixtures of various chiral compounds (ie, racemates). In all compounds of this application, each chiral carbon atom can optionally be in the R configuration or the S configuration, or a mixture of the R configuration and the S configuration.
如本文所用，术语“本申请化合物”指本申请的化合物。该术语还包括本申请化合物的各种晶型形式、药学上可接受的盐、水合物或溶剂合物。As used herein, the term "compound of the present application" refers to a compound of the present application. The term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of the present application.
当本文中使用商品名时，该商品名意在包括商品名产品制剂、其相应的仿制药，以及商品名产品的活性药物组分。When a tradename is used herein, the tradename is intended to include the tradename product formulation, its corresponding generic drug, and the active pharmaceutical ingredient of the tradename product.
如本文所用，“抗体”以其最广泛的含义使用并且特别覆盖单克隆抗体、多克隆抗体、二聚体、多聚体、多特异性抗体(例如双特异性抗体)和抗体片段，只要它们表现出所需的生物活性。抗体可以为鼠、人、人源化、嵌合的抗体或来源于其它物种。抗体为由能够识别和结合特异性抗原的免疫系统产生的蛋白质。靶抗原一般具有由多种抗体的CDRs识别的大量结合位点，也称作表位。特异性结合不同表位的各抗体具有不同的结构。因此，一种抗原可以 具有一种以上相应的抗体。抗体包括全-长免疫球蛋白分子或全-长免疫球蛋白分子的免疫活性部分，即含有特异性结合所关注靶标的抗原或其部分的分子，这类靶标包括，但不限于癌细胞或产生与自身免疫性疾病相关的自身免疫抗体的细胞。本申请描述的免疫球蛋白可以具有免疫球蛋白分子的任意类型(例如IgG、IgE、IgM、IgD和IgA)、类别(例如IgG1、IgG2、IgG3、IgG4、IgA1和IgA2)或亚类。免疫球蛋白可以来源于任意的物种。然而，在一个方面中，免疫球蛋白来源于人、鼠或兔。“抗体片段”可以包含全长抗体的一部分，一般为其抗原结合区或可变区。抗体片段的实例包括：Fab、Fab’、F(ab’)2和Fv片段；双抗体；线性抗体；微抗体(minibody)；Fab表达文库制备的片段；抗-独特型(抗-Id)抗体；CDR(互补决定区)；和以免疫特异性方式结合癌细胞抗原、病毒抗原或微生物抗原的上述任意的表位-结合片段；单链抗体分子；和由抗体片段形成的多特异性抗体。本申请中组成抗体药物偶联物的抗体可以保持其原有野生状态时的抗原结合能力。因此，本申请中的抗体可以，例如可以专一性地，与抗原结合。涉及的抗原包括，例如，肿瘤相关抗原(TAA)，细胞表面受体蛋白和其他细胞表面分子，细胞存活调节因子，细胞增殖调节因子，与组织生长与分化相关的分子(如已知或预知的具有功能性的)，淋巴因子，细胞因子，参与细胞循环调节的分子，参与血管生成的分子，以及与血管生成有关的分子(如已知抗体结合的抗原可以是上述分类中一个或一个子集，而其它的子集则包含其它的具有特殊性质的分子/抗原(与目标抗原相比)。应用在抗体药物偶联物中的抗体包括，但不局限于，针对细胞表面受体和肿瘤相关抗原的抗体。这样的肿瘤相关抗原是业内所熟知的，可以通过业内熟知的抗体制备方法和信息来制备。这些目标物能够特异性地表达在一种或多种癌症细胞表面，而在一种或多种非癌细胞表面表达很少或不表达。通常，相对于非癌细胞表面而言,这样的肿瘤相关多肽在癌细胞表面可以更加过度表达。As used herein, "antibody" is used in its broadest sense and specifically covers monoclonal antibodies, polyclonal antibodies, dimers, multimers, multispecific antibodies (eg, bispecific antibodies), and antibody fragments, so long as they are exhibit the desired biological activity. Antibodies can be murine, human, humanized, chimeric, or derived from other species. Antibodies are proteins produced by the immune system capable of recognizing and binding specific antigens. Target antigens generally have a large number of binding sites, also referred to as epitopes, recognized by the CDRs of various antibodies. Each antibody that specifically binds to a different epitope has a different structure. Thus, an antigen can have more than one corresponding antibody. Antibodies include full-length immunoglobulin molecules or immunologically active portions of full-length immunoglobulin molecules, i.e., molecules that contain an antigen or portion thereof that specifically binds a target of interest, including, but not limited to, cancer cells or Cells with autoimmune antibodies associated with autoimmune diseases. The immunoglobulins described herein can be of any class (eg, IgG, IgE, IgM, IgD, and IgA), class (eg, IgGl, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass of immunoglobulin molecules. Immunoglobulins can be derived from any species. However, in one aspect, the immunoglobulin is derived from human, murine or rabbit. An "antibody fragment" may comprise a portion of a full-length antibody, typically its antigen-binding or variable region. Examples of antibody fragments include: Fab, Fab', F(ab')2 and Fv fragments; diabodies; linear antibodies; minibodies; Fab expression library prepared fragments; anti-idiotypic (anti-Id) antibodies ; CDRs (complementarity determining regions); and any of the above epitope-binding fragments that immunospecifically bind to cancer cell antigens, viral antigens, or microbial antigens; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. The antibody constituting the antibody-drug conjugate in the present application can maintain the antigen-binding ability of its original wild state. Thus, the antibodies of the present application can, for example, specifically bind to an antigen. Antigens involved include, for example, tumor-associated antigens (TAAs), cell surface receptor proteins and other cell surface molecules, regulators of cell survival, regulators of cell proliferation, molecules associated with tissue growth and differentiation (as known or predicted) functional), lymphokines, cytokines, molecules involved in the regulation of the cell cycle, molecules involved in angiogenesis, and molecules involved in angiogenesis (eg known antibodies bind antigens can be one or a subset of the above categories , while other subsets contain other molecules/antigens with specific properties (compared to the target antigen). Antibodies used in antibody drug conjugates include, but are not limited to, targeting cell surface receptors and tumor-associated Antibodies to antigens. Such tumor-associated antigens are well known in the industry and can be prepared by well-known antibody preparation methods and information in the industry. These targets can be specifically expressed on the surface of one or more cancer cells, while in a or a variety of non-cancerous cell surfaces expressed little or no. Generally, such tumor-associated polypeptides can be more overexpressed on the surface of cancer cells relative to the surface of non-cancer cells.
在本申请中，术语“恩诺单抗”通常是指一种靶向Nectin-4的抗体。例如，恩诺单抗(Enfortumab)可以记载在WO2017042210A1中。在本申请中，恩诺单抗可以是指任何一种包含恩诺单抗的重链可变区CDR1-3和轻链可变区CDR1-3的抗体或抗原结合片段。在本申请中，恩诺单抗可以是指任何一种包含恩诺单抗的重链可变区和轻链可变区的抗体或抗原结合片段。In this application, the term "enrozumab" generally refers to an antibody that targets Nectin-4. For example, Enfortumab can be described in WO2017042210A1. In the present application, ennozumab may refer to any antibody or antigen-binding fragment comprising the heavy chain variable region CDR1-3 and light chain variable region CDR1-3 of enrozumab. In the present application, ennozumab may refer to any antibody or antigen-binding fragment comprising the heavy chain variable region and light chain variable region of enrozumab.
在本申请中，术语“帕妥珠单抗”通常是指一种靶向HER2的抗体。例如，帕妥珠单抗(Pertuzumab)可以记载在WO2014172371A2中。在本申请中，帕妥珠单抗可以是指任何一种包含帕妥珠单抗的重链可变区CDR1-3和轻链可变区CDR1-3的抗体或抗原结合片段。在 本申请中，帕妥珠单抗可以是指任何一种包含帕妥珠单抗的重链可变区和轻链可变区的抗体或抗原结合片段。In this application, the term "Pertuzumab" generally refers to an antibody that targets HER2. For example, Pertuzumab can be described in WO2014172371A2. In the present application, Pertuzumab may refer to any antibody or antigen-binding fragment comprising the heavy chain variable region CDR1-3 and the light chain variable region CDR1-3 of Pertuzumab. In the present application, Pertuzumab can refer to any antibody or antigen-binding fragment comprising the variable region of the heavy chain and the variable region of the light chain of Pertuzumab.
在本申请中，术语“曲妥珠单抗”通常是指一种靶向HER2的抗体。例如，曲妥珠单抗(Trastuzumab)可以记载在US20060275305A1中。在本申请中，曲妥珠单抗可以是指任何一种包含曲妥珠单抗的重链可变区CDR1-3和轻链可变区CDR1-3的抗体或抗原结合片段。在本申请中，曲妥珠单抗可以是指任何一种包含曲妥珠单抗的重链可变区和轻链可变区的抗体或抗原结合片段。In this application, the term "trastuzumab" generally refers to an antibody that targets HER2. For example, Trastuzumab can be described in US20060275305A1. In the present application, trastuzumab may refer to any antibody or antigen-binding fragment comprising the heavy chain variable region CDR1-3 and light chain variable region CDR1-3 of trastuzumab. In the present application, trastuzumab may refer to any antibody or antigen-binding fragment comprising the heavy and light chain variable regions of trastuzumab.
在本申请中，术语“赛妥珠单抗”通常是指一种靶向TROP2的抗体。例如，赛妥珠单抗(Sacituzumab，hRS7)可以记载在WO2003074566中。在本申请中，赛妥珠单抗可以是指任何一种包含赛妥珠单抗的重链可变区CDR1-3和轻链可变区CDR1-3的抗体或抗原结合片段。在本申请中，赛妥珠单抗可以是指任何一种包含赛妥珠单抗的重链可变区和轻链可变区的抗体或抗原结合片段。In this application, the term "certolizumab" generally refers to an antibody that targets TROP2. For example, Sacituzumab (hRS7) can be described in WO2003074566. In the present application, certolizumab may refer to any antibody or antigen-binding fragment comprising the heavy chain variable region CDR1-3 and light chain variable region CDR1-3 of certolizumab. In the present application, certolizumab may refer to any antibody or antigen-binding fragment comprising the heavy and light chain variable regions of certolizumab.
在本申请中，术语“Patritumab”通常是指一种靶向HER3的抗体。例如，Patritumab可以记载在CN102174105B中。在本申请中，Patritumab单抗可以是指任何一种包含Patritumab的重链可变区CDR1-3和轻链可变区CDR1-3的抗体或抗原结合片段。在本申请中，Patritumab可以是指任何一种包含Patritumab的重链可变区和轻链可变区的抗体或抗原结合片段。In this application, the term "Patritumab" generally refers to an antibody that targets HER3. For example, Patritumab can be described in CN102174105B. In the present application, Patritumab monoclonal antibody may refer to any antibody or antigen-binding fragment comprising the heavy chain variable region CDR1-3 and light chain variable region CDR1-3 of Patritumab. In the present application, Patritumab may refer to any antibody or antigen-binding fragment comprising the variable region of the heavy chain and the variable region of the light chain of Patritumab.
在本申请中，术语“抗体H01L02”通常是指一种靶向CDH6的抗体。例如，单克隆抗体H01L02可以记载在WO2018212136、US20200171163A1中。例如，H01L02单抗可以是药物DS6000所采用的单抗。在本申请中，H01L02单抗可以是指任何一种包含H01L02单抗的重链可变区CDR1-3和轻链可变区CDR1-3的抗体或抗原结合片段。在本申请中，H01L02单抗可以是指任何一种包含H01L02单抗的重链可变区和轻链可变区的抗体或抗原结合片段。In this application, the term "antibody H01L02" generally refers to an antibody targeting CDH6. For example, the monoclonal antibody H01L02 can be described in WO2018212136, US20200171163A1. For example, the H01L02 mAb can be the mAb used by the drug DS6000. In the present application, the H01L02 monoclonal antibody may refer to any antibody or antigen-binding fragment comprising the heavy chain variable region CDR1-3 and the light chain variable region CDR1-3 of the H01L02 monoclonal antibody. In the present application, the H01L02 mAb can refer to any antibody or antigen-binding fragment comprising the heavy chain variable region and the light chain variable region of the H01L02 mAb.
在本申请中，术语“多肽残基”通常是指包含一个或多个氨基酸残基连接而成的残基。例如，多肽残基中的一个或多个氨基酸可以是任选取代的。例如，本申请的多肽残基可以选自以下组：苯丙氨酸-赖氨酸(Phe-Lys)、缬氨酸-丙氨酸(Val-Ala)、缬氨酸-瓜氨酸(Val-Cit)、谷氨酸-缬氨酸-丙氨酸(Glu-Val-Ala)、谷氨酸-缬氨酸-瓜氨酸(Glu-Val-Cit)、缬氨酸-赖氨酸(Val-Lys)、丙氨酸-丙氨酸-丙氨酸(Ala-Ala-Ala)、丙氨酸-丙氨酸-天冬酰胺(Ala-Ala-Asn)和甘氨酸-甘氨酸-苯丙氨酸-甘氨酸(Gly-Gly-Phe-Gly)。In this application, the term "polypeptide residue" generally refers to a residue comprising one or more amino acid residues linked together. For example, one or more amino acids in a polypeptide residue may be optionally substituted. For example, the polypeptide residues of the present application may be selected from the group consisting of phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala), valine-citrulline (Val) -Cit), glutamic acid-valine-alanine (Glu-Val-Ala), glutamic acid-valine-citrulline (Glu-Val-Cit), valine-lysine ( Val-Lys), Alanine-Alanine-Alanine (Ala-Ala-Ala), Alanine-Alanine-Asparagine (Ala-Ala-Asn), and Glycine-Glycine-Phenylalanine Acid-glycine (Gly-Gly-Phe-Gly).
在本申请中，术语“聚乙二醇基”通常是指包含一个或多个乙二醇残基连接而成的残基。例如，聚乙二醇基可以包含-(CH 2CH 2O) p-，其中p为至少为1的数。例如本申请中的聚乙二醇基可以是任选取代的。 In this application, the term "polyethylene glycol" generally refers to a residue comprising one or more ethylene glycol residues linked together. For example, a polyethylene glycol group may comprise - ( CH2CH2O ) p- , where p is a number of at least one. For example, polyethylene glycol groups in this application may be optionally substituted.
在本申请中，术语“甘醇基”通常是指聚乙二醇基。例如本申请中的甘醇基可以是任选取代的。例如，甘醇基之前的数字可以表示甘醇基的乙二醇单元的数量，例如二甘醇基可以是指两个乙二醇聚合的残基。In this application, the term "glycol group" generally refers to a polyethylene glycol group. For example, glycol groups in this application may be optionally substituted. For example, a number preceding a glycol group may indicate the number of ethylene glycol units of a glycol group, eg, a diethylene glycol group may refer to the polymerized residue of two glycols.
在本申请中，术语“聚肌氨酸残基”通常是指包含一个或多个肌氨酸残基连接而成的残基。例如，聚肌氨酸残基可以包含-(COCH 2N(CH 3)) q-，其中q为至少为1的数。例如本申请中的聚肌氨酸残基可以是任选取代的。例如，包含聚肌氨酸残基的结构可以是
Figure PCTCN2022089726-appb-000005
其中n2为4至18的数。 In this application, the term "polysarcosine residue" generally refers to a residue comprising one or more sarcosine residues linked together. For example, a polysarcosine residue may comprise -(COCH2N( CH3 ) ) q- , where q is a number of at least one. For example, polysarcosine residues in the present application may be optionally substituted. For example, a structure containing polysarcosine residues can be
Figure PCTCN2022089726-appb-000005
where n2 is a number from 4 to 18.
在本申请中，术语“十二烷基硫酸钠聚丙烯酰胺凝胶电泳”通常是指一种物质分析表征技术。例如十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)可以检测物质分子量的大小。In this application, the term "sodium dodecyl sulfate polyacrylamide gel electrophoresis" generally refers to an analytical characterization technique for substances. For example, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) can detect the molecular weight of substances.
在本申请中，术语“疏水层析”通常是指一种基于的是物质疏水差异的分析技术。In this application, the term "hydrophobic chromatography" generally refers to an analytical technique based on differences in the hydrophobicity of substances.
在本申请中，术语“液相质谱”通常是指一种鉴定物质组分的分析方法。例如液相质谱可以通过液相色谱-质谱联用分析待测物质的分子量。In this application, the term "liquid phase mass spectrometry" generally refers to an analytical method for identifying constituents of matter. For example, liquid chromatography-mass spectrometry can analyze the molecular weight of the substance to be tested by liquid chromatography-mass spectrometry.
在本申请中，术语“肿瘤”通常是指任何新的病理性的组织增生。对于本申请来说，血管生成是肿瘤特征的一部分。肿瘤可能是良性的，也可能是恶性的。术语“肿瘤”一般用于指良性或恶性的肿瘤，而术语“癌”一般用于指恶性肿瘤，可以是转移癌，也可以是非转移癌。可用本申请的方法诊断的肿瘤选自以下组：乳腺癌、卵巢癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、急性淋巴细胞性白血病、间变性大细胞淋巴瘤、多发性骨髓瘤、前列腺癌、非小细胞肺癌、小细胞肺癌、恶性黑色素瘤、鳞状细胞癌、胶质母细胞瘤、肾细胞癌、胃肠道肿瘤、胰腺癌、前列腺癌、直结肠、胃癌、神经胶质瘤和间皮瘤。用于研究时，可通过本领域技术人员熟知的方法从易于获得的资源中将这些组织分离出来。In this application, the term "tumor" generally refers to any new pathological tissue proliferation. For the purposes of this application, angiogenesis is part of the tumor profile. Tumors can be benign or malignant. The term "tumor" is generally used to refer to benign or malignant tumors, while the term "cancer" is generally used to refer to malignant tumors, which may be metastatic or non-metastatic. Tumors diagnosable by the methods of the present application are selected from the group consisting of breast cancer, ovarian cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, acute lymphoblastic leukemia, anaplastic large cell lymphoma, multiple myeloma, Prostate cancer, non-small cell lung cancer, small cell lung cancer, malignant melanoma, squamous cell carcinoma, glioblastoma, renal cell carcinoma, gastrointestinal tumors, pancreatic cancer, prostate cancer, rectal colon, gastric cancer, glial tumor and mesothelioma. For research, these tissues can be isolated from readily available sources by methods well known to those skilled in the art.
发明详述Detailed description of the invention
一方面，本申请提供一种化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，其中所述化合物可以包含式(C-Trop-2)所示的结构：In one aspect, the application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable compound thereof. An acceptable salt, prodrug or solvate, wherein the compound may comprise the structure of formula (C-Trop-2):
Figure PCTCN2022089726-appb-000006
Figure PCTCN2022089726-appb-000006
其中，Q 1可以包含连接体， where Q 1 may contain a linker,
L 1可以包含-L 1a-C(＝O)-， L 1 may contain -L 1a -C(=O)-,
其中，L 1a可以选自以下组：任选取代的亚烷基、任选取代的聚乙二醇基、任选取代的亚烯基、任选取代的亚炔基、任选取代的亚脂环基、任选取代的亚脂杂环基、任选取代的亚芳基和任选取代的亚杂芳基； Wherein, L 1a can be selected from the following group: optionally substituted alkylene, optionally substituted polyethylene glycol, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted idene cyclyl, optionally substituted alicyclic heterocyclyl, optionally substituted arylene, and optionally substituted heteroarylene;
L 2可以包含任选取代的多肽残基， L2 may comprise optionally substituted polypeptide residues,
L 3可以包含任选取代的间隔基团，例如，本申请的间隔基团可以是具有自降解能力。例如本申请的间隔基团可以包含任选取代的
Figure PCTCN2022089726-appb-000007
或任选取代的
Figure PCTCN2022089726-appb-000008
L 3 may contain an optionally substituted spacer group, for example, the spacer group of the present application may be self-degradable. For example, the spacer groups of the present application may comprise optionally substituted
Figure PCTCN2022089726-appb-000007
or optionally substituted
Figure PCTCN2022089726-appb-000008
其中，L 2和/或L 3可以包含任选取代的聚肌氨酸残基， wherein L2 and/or L3 may comprise optionally substituted polysarcosine residues,
T可以包含药物单元，T may contain drug units,
Ab可以为能够结合Trop-2的配体，m可以为1-8的数。Ab can be a ligand capable of binding Trop-2, and m can be a number from 1-8.
另一方面，本申请还提供一种化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，其中所述化合物可以包含式(C-Trop-2-M)所示的结构：In another aspect, the present application also provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or a mixture thereof A pharmaceutically acceptable salt, prodrug or solvate, wherein the compound may comprise a structure represented by the formula (C-Trop-2-M):
Figure PCTCN2022089726-appb-000009
Figure PCTCN2022089726-appb-000009
其中，Q 1可以包含连接体， where Q 1 may contain a linker,
L 1可以包含-L 1a-C(＝O)-， L 1 may contain -L 1a -C(=O)-,
其中，L 1a可以选自以下组：任选取代的亚烷基、任选取代的聚乙二醇基、任选取代的亚烯基、任选取代的亚炔基、任选取代的亚脂环基、任选取代的亚脂杂环基、任选取代的亚芳基和任选取代的亚杂芳基； Wherein, L 1a can be selected from the following group: optionally substituted alkylene, optionally substituted polyethylene glycol, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted idene cyclyl, optionally substituted alicyclic heterocyclyl, optionally substituted arylene, and optionally substituted heteroarylene;
L 2可以包含任选取代的多肽残基， L2 may comprise optionally substituted polypeptide residues,
L 3可以包含任选取代的间隔基团，例如，本申请的间隔基团可以是具有自降解能力。例如本申请的间隔基团可以包含任选取代的
Figure PCTCN2022089726-appb-000010
或任选取代的
Figure PCTCN2022089726-appb-000011
L 3 may contain an optionally substituted spacer group, for example, the spacer group of the present application may be self-degradable. For example, the spacer groups of the present application may comprise optionally substituted
Figure PCTCN2022089726-appb-000010
or optionally substituted
Figure PCTCN2022089726-appb-000011
其中，L 2和/或L 3可以包含任选取代的结构单元-X， Wherein, L 2 and/or L 3 may comprise optionally substituted structural unit-X,
T可以包含药物单元，T may contain drug units,
Ab可以为能够结合Trop-2的配体，m可以为1-8的数。Ab can be a ligand capable of binding Trop-2, and m can be a number from 1-8.
例如，其中，L 3选自以下组：任选取代的
Figure PCTCN2022089726-appb-000012
和任选取代的
Figure PCTCN2022089726-appb-000013
For example, wherein L is selected from the group of: optionally substituted
Figure PCTCN2022089726-appb-000012
and optionally substituted
Figure PCTCN2022089726-appb-000013
例如，其中，L 3的苯环可以被任选取代的结构单元-X取代。例如，该结构单元-X可以选自以下组：任选取代的
Figure PCTCN2022089726-appb-000014
其中X 1选自以下组：羰基、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；其中X 2选自以下组：氢、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；其中X 3选自以下组：氢、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；所述的C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基各自独立地任选地被选自氘代、卤素、氰基、硝基、氨基、烷基、羧基、烷氧基或环烷基的一个或者多个取代基取代。 For example, wherein, the benzene ring of L 3 may be substituted by the optionally substituted structural unit -X. For example, the structural unit -X can be selected from the group of: optionally substituted
Figure PCTCN2022089726-appb-000014
wherein X 1 is selected from the group consisting of carbonyl, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight chain heteroalkyl containing 1-8 atoms, and Linear-cyclic heteroalkyl groups containing 1-8 atoms containing 1-3 atoms selected from N, O or S ; wherein X is selected from the group consisting of hydrogen, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkane containing 1-8 atoms group, the heteroalkyl group contains 1-3 atoms selected from N, O or S; wherein X 3 is selected from the following group: hydrogen, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 - C6 cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkyl containing 1-8 atoms, the heteroalkyl containing selected from N, O or 1-3 atoms of S; the C 1 -C 8 alkyl group, the C 1 -C 8 alkoxy group, the C 1 -C 6 cycloalkyl group, the straight chain heteroalkyl group containing 1-8 atoms, and straight-chain-cyclic heteroalkyl groups containing 1-8 atoms each independently optionally selected from deuterated, halogen, cyano, nitro, amino, alkyl, carboxy, alkoxy, or cycloalkyl substituted with one or more substituents.
例如，其中，L 3的苯环可以被任选取代的结构单元-X取代。例如，该结构单元-X可以包含任选取代的
Figure PCTCN2022089726-appb-000015
其中X 1选自以下组：C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子，所述的C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基各自独立地任选地选自氘代、卤素、氰基、硝基、氨基、烷基、羧基、烷氧基或环烷基的一个或者多个取代基取代。 For example, wherein, the benzene ring of L 3 may be substituted by the optionally substituted structural unit -X. For example, the building block -X may contain an optionally substituted
Figure PCTCN2022089726-appb-000015
wherein X 1 is selected from the group consisting of C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight chain heteroalkyl containing 1-8 atoms, and containing 1 -8-atom straight-chain-cyclic heteroalkyl, said heteroalkyl containing 1-3 atoms selected from N, O or S, said C 1 -C 8 alkyl, C 1 -C 8 Alkoxy, C1 - C6cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkyl containing 1-8 atoms are each independently optionally selected from Substituted with one or more substituents of deuterated, halogen, cyano, nitro, amino, alkyl, carboxyl, alkoxy or cycloalkyl.
例如，其中，结构单元-X为任选取代的
Figure PCTCN2022089726-appb-000016
For example, wherein the structural unit -X is optionally substituted
Figure PCTCN2022089726-appb-000016
例如，本申请提供一种化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，其中所述化合物可以包含式(C-Trop-2-M2)所示的结构：For example, the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C-Trop-2-M2):
Figure PCTCN2022089726-appb-000017
Figure PCTCN2022089726-appb-000017
其中，L可以含任选取代的亚烷基、任选取代的聚乙二醇基和任选取代的亚脂环基；R 1可以为任选取代的异丙基或任选取代的苄基；R 2可以为任选取代的甲基、任选取代的
Figure PCTCN2022089726-appb-000018
或任选取代的
Figure PCTCN2022089726-appb-000019
R 3可以为氢或R 5；其中，R 5可以为任选取代的
Figure PCTCN2022089726-appb-000020
T可以包含伊沙替康(Exatecan，CAS登录号171335-80-1)和/或贝洛替康(Belotecan，CAS登录号256411-32-2)和/或Genz-644282(CAS登录号529488-28-6)。 Wherein, L can contain an optionally substituted alkylene group, an optionally substituted polyethylene glycol group and an optionally substituted alicyclic group; R 1 can be an optionally substituted isopropyl group or an optionally substituted benzyl group ; R 2 can be optionally substituted methyl, optionally substituted
Figure PCTCN2022089726-appb-000018
or optionally substituted
Figure PCTCN2022089726-appb-000019
R 3 can be hydrogen or R 5 ; wherein R 5 can be optionally substituted
Figure PCTCN2022089726-appb-000020
T may comprise ixatecan (Exatecan, CAS accession number 171335-80-1) and/or belonotecan (Belotecan, CAS accession number 256411-32-2) and/or Genz-644282 (CAS accession number 529488- 28-6).
例如，其中，Q 1可以包含与巯基偶联后的连接体。 For example, Q 1 may comprise a linker coupled with a thiol group.
例如，其中，Q 1可以选自以下组：任选取代的
Figure PCTCN2022089726-appb-000021
任选取代的
Figure PCTCN2022089726-appb-000022
任选取代的
Figure PCTCN2022089726-appb-000023
和任选取代的
Figure PCTCN2022089726-appb-000024
For example, wherein Q 1 can be selected from the group of: optionally substituted
Figure PCTCN2022089726-appb-000021
optionally substituted
Figure PCTCN2022089726-appb-000022
optionally substituted
Figure PCTCN2022089726-appb-000023
and optionally substituted
Figure PCTCN2022089726-appb-000024
例如，其中，L 1a可以选自以下组：任选取代的C 1-C 7的亚烷基、任选取代的二甘醇基至八甘醇基、任选取代的C 3-C 6亚脂环基、任选取代的亚芳基和任选取代的亚杂芳基。 For example, wherein, L 1a may be selected from the group consisting of optionally substituted C 1 -C 7 alkylene, optionally substituted diethylene glycol to octaethylene glycol, optionally substituted C 3 -C 6 alkylene Alicyclic, optionally substituted arylene, and optionally substituted heteroarylene.
例如，其中，L 1a可以选自以下组：任选取代的亚甲基、任选取代的亚乙基、任选取代的亚丙基、任选取代的亚丁基、任选取代的亚戊基、任选取代的二甘醇基、任选取代的四甘醇基、任选取代的六甘醇基、任选取代的八甘醇基和任选取代的亚环己基。 For example, wherein L 1a may be selected from the group consisting of optionally substituted methylene, optionally substituted ethylene, optionally substituted propylene, optionally substituted butylene, optionally substituted pentylene , optionally substituted diethylene glycol, optionally substituted tetraethylene glycol, optionally substituted hexaethylene glycol, optionally substituted octaethylene glycol, and optionally substituted cyclohexylene.
例如，其中，L 1a可以选自以下组：任选取代的亚甲基、任选取代的亚乙基、任选取代的亚丙基、任选取代的亚丁基、和任选取代的亚戊基。 For example, wherein L 1a can be selected from the group consisting of optionally substituted methylene, optionally substituted ethylene, optionally substituted propylene, optionally substituted butylene, and optionally substituted pentylene base.
例如，其中，L 1a可以选自以下组：任选取代的二甘醇基、任选取代的三甘醇基、任选取代的四甘醇基、任选取代的五甘醇基、任选取代的六甘醇基、任选取代的七甘醇基、和任选取代的八甘醇基。 For example, wherein, L 1a can be selected from the group of: optionally substituted diethylene glycol, optionally substituted triethylene glycol, optionally substituted tetraethylene glycol, optionally substituted pentaethylene glycol, optionally substituted Substituted hexaethylene glycol, optionally substituted heptaethylene glycol, and optionally substituted octaethylene glycol.
例如，其中，L 1a可以选自以下组：任选取代的亚环丙基、任选取代的亚环丁基和任选取代的亚环己基。 For example, wherein L 1a may be selected from the group consisting of optionally substituted cyclopropylene, optionally substituted cyclobutylene, and optionally substituted cyclohexylene.
例如，其中，L 2可以包含任选取代的选自以下组的氨基酸构成的多肽残基：苯丙氨酸、异亮氨酸、亮氨酸、色氨酸、缬氨酸、甲硫氨酸、酪氨酸、丙氨酸、苏氨酸、组氨酸、丝氨 酸、谷氨酰胺、精氨酸、赖氨酸、天冬酰胺、谷氨酸、脯氨酸、瓜氨酸、天冬氨酸和甘氨酸。 For example, wherein L2 may comprise optionally substituted polypeptide residues consisting of amino acids selected from the group consisting of phenylalanine, isoleucine, leucine, tryptophan, valine, methionine , tyrosine, alanine, threonine, histidine, serine, glutamine, arginine, lysine, asparagine, glutamic acid, proline, citrulline, aspartate acid and glycine.
例如，其中，L 2可以包含任选取代的选自以下组的氨基酸构成的多肽残基：甘氨酸、苯丙氨酸、缬氨酸、丙氨酸、精氨酸、瓜氨酸、天冬氨酸、天冬酰胺和赖氨酸。 For example, wherein L 2 may comprise optionally substituted polypeptide residues consisting of amino acids selected from the group consisting of glycine, phenylalanine, valine, alanine, arginine, citrulline, aspartate acid, asparagine and lysine.
例如，其中，L 2可以包含任选取代的选自以下组的多肽残基：苯丙氨酸-赖氨酸(Phe-Lys)、缬氨酸-丙氨酸(Val-Ala)、缬氨酸-瓜氨酸(Val-Cit)、谷氨酸-缬氨酸-丙氨酸(Glu-Val-Ala)、谷氨酸-缬氨酸-瓜氨酸(Glu-Val-Cit)、缬氨酸-赖氨酸(Val-Lys)、丙氨酸-丙氨酸-丙氨酸(Ala-Ala-Ala)、丙氨酸-丙氨酸-天冬酰胺(Ala-Ala-Asn)和甘氨酸-甘氨酸-苯丙氨酸-甘氨酸(Gly-Gly-Phe-Gly)。 For example, wherein L 2 may comprise an optionally substituted polypeptide residue selected from the group consisting of: phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala), valine Acid-citrulline (Val-Cit), glutamic acid-valine-alanine (Glu-Val-Ala), glutamic acid-valine-citrulline (Glu-Val-Cit), valine Amino-Lysine (Val-Lys), Alanine-Alanine-Alanine (Ala-Ala-Ala), Alanine-Alanine-Asparagine (Ala-Ala-Asn) and Glycine-Glycine-Phenylalanine-Glycine (Gly-Gly-Phe-Gly).
例如，其中，L 2可以包含任选取代的选自以下组的多肽残基：苯丙氨酸-赖氨酸(Phe-Lys)、缬氨酸-丙氨酸(Val-Ala)、缬氨酸-瓜氨酸(Val-Cit)和缬氨酸-赖氨酸(Val-Lys)。 For example, wherein L 2 may comprise an optionally substituted polypeptide residue selected from the group consisting of: phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala), valine Acid-citrulline (Val-Cit) and valine-lysine (Val-Lys).
例如，其中，当L 2包含赖氨酸残基时，所述赖氨酸残基可以被包含聚肌氨酸残基的结构R 1取代。 For example, where L2 comprises a lysine residue, the lysine residue can be substituted by a structure R1 comprising a polysarcosine residue.
例如，L 2包含的任意的H可以被R 1取代。 For example, any H contained in L 2 may be substituted by R 1 .
例如，其中，所述R 1可以为任选取代的
Figure PCTCN2022089726-appb-000025
其中n1为4至18的数，R选自以下组：C 1-C 6烷基、C 1-C 6环烷基、和C 1-C 6烷氧基。 For example, wherein, the R 1 can be optionally substituted
Figure PCTCN2022089726-appb-000025
wherein n1 is a number from 4 to 18, and R is selected from the group consisting of C 1 -C 6 alkyl, C 1 -C 6 cycloalkyl, and C 1 -C 6 alkoxy.
例如，其中，n1可以为4至18、8至18、4至12或8至12。例如，其中，n1可以为4、5、6、7、8、9、10、11、12、13、14、15、16、17或18。For example, where n1 can be 4 to 18, 8 to 18, 4 to 12, or 8 to 12. For example, where n1 can be 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18.
例如，其中，L 3选自以下组：任选取代的
Figure PCTCN2022089726-appb-000026
和任选取代的
Figure PCTCN2022089726-appb-000027
For example, wherein L is selected from the group of: optionally substituted
Figure PCTCN2022089726-appb-000026
and optionally substituted
Figure PCTCN2022089726-appb-000027
例如，其中，L 3的苯环可以被包含聚肌氨酸残基的结构R 2取代。 For example, where the benzene ring of L3 can be substituted with a structure R2 comprising a polysarcosine residue.
例如，L 3的苯环包含的任意的H可以被R 2取代。 For example, any H contained in the benzene ring of L 3 may be substituted by R 2 .
例如，其中，L 3的苯环与任选取代的聚肌氨酸残基通过结构单元-X-连接，结构单元-X-选自以下组：任选取代的
Figure PCTCN2022089726-appb-000028
其中X 1选自以下组：羰基、C 1-C 8烷基、C 1-C 8烷氧 基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；其中X 2选自以下组：氢、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；其中X 3为共价键或选自以下组：氢、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；所述的C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基各自独立地任选地被选自氘代、卤素、氰基、硝基、氨基、烷基、羧基、烷氧基或环烷基的一个或者多个取代基取代。 For example, wherein the benzene ring of L 3 is connected to the optionally substituted polysarcosine residue through the structural unit -X-, the structural unit -X- is selected from the group consisting of: optionally substituted
Figure PCTCN2022089726-appb-000028
wherein X 1 is selected from the group consisting of carbonyl, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight chain heteroalkyl containing 1-8 atoms, and Linear-cyclic heteroalkyl groups containing 1-8 atoms containing 1-3 atoms selected from N, O or S ; wherein X is selected from the group consisting of hydrogen, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkane containing 1-8 atoms group, the heteroalkyl group contains 1-3 atoms selected from N, O or S; wherein X 3 is a covalent bond or is selected from the following group: hydrogen, C 1 -C 8 alkyl, C 1 -C 8 Alkoxy, C1 - C6cycloalkyl, straight-chain heteroalkyl containing 1-8 atoms, and straight-cyclic-cyclic heteroalkyl containing 1-8 atoms, the heteroalkyl containing selected 1-3 atoms from N, O or S; said C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight containing 1-8 atoms Chain heteroalkyl, and straight-chain-cyclic heteroalkyl containing 1-8 atoms are each independently optionally selected from deuterated, halogen, cyano, nitro, amino, alkyl, carboxy, alkoxy substituted with one or more substituents of the radical or cycloalkyl.
例如，L 3的苯环与任选取代的聚肌氨酸残基通过结构单元-X-连接，结构单元-X-选自，但不局限于：
Figure PCTCN2022089726-appb-000029
其中X 1选自以下组：C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子，所述的C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基各自独立地任选地选自氘代、卤素、氰基、硝基、氨基、烷基、羧基、烷氧基或环烷基的一个或者多个取代基取代。 For example, the benzene ring of L 3 is connected to the optionally substituted polysarcosine residue through a structural unit -X-, which is selected from, but is not limited to:
Figure PCTCN2022089726-appb-000029
wherein X 1 is selected from the group consisting of C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight chain heteroalkyl containing 1-8 atoms, and containing 1 -8-atom straight-chain-cyclic heteroalkyl, said heteroalkyl containing 1-3 atoms selected from N, O or S, said C 1 -C 8 alkyl, C 1 -C 8 Alkoxy, C1 - C6cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkyl containing 1-8 atoms are each independently optionally selected from Substituted with one or more substituents of deuterated, halogen, cyano, nitro, amino, alkyl, carboxyl, alkoxy or cycloalkyl.
例如，其中，结构单元-X-为任选取代的
Figure PCTCN2022089726-appb-000030
所述任选取代的聚肌氨酸残基包含
Figure PCTCN2022089726-appb-000031
其中n2为4至18的数，R选自以下组：C 1-C 6烷基、C 1-C 6环烷基、和C 1-C 6烷氧基。 For example, wherein the structural unit -X- is optionally substituted
Figure PCTCN2022089726-appb-000030
The optionally substituted polysarcosine residue comprises
Figure PCTCN2022089726-appb-000031
wherein n2 is a number from 4 to 18, and R is selected from the group consisting of C 1 -C 6 alkyl, C 1 -C 6 cycloalkyl, and C 1 -C 6 alkoxy.
例如，其中，结构单元-X-选自以下组：任选取代的
Figure PCTCN2022089726-appb-000032
其中X 1选自以下组：羰基、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；其中X 2选自以下组：氢、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；其中X 3为共价键或选自以下组：氢、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O 或者S的1-3个原子；所述的C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基各自独立地任选地被选自氘代、卤素、氰基、硝基、氨基、烷基、羧基、烷氧基或环烷基的一个或者多个取代基取代，所述任选取代的聚肌氨酸残基包含
Figure PCTCN2022089726-appb-000033
其中n2为4至18的数，R选自以下组：C 1-C 6烷基、C 1-C 6环烷基、和C 1-C 6烷氧基。 For example, wherein the structural unit -X- is selected from the group consisting of optionally substituted
Figure PCTCN2022089726-appb-000032
wherein X 1 is selected from the group consisting of carbonyl, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight chain heteroalkyl containing 1-8 atoms, and Linear-cyclic heteroalkyl groups containing 1-8 atoms containing 1-3 atoms selected from N, O or S ; wherein X is selected from the group consisting of hydrogen, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkane containing 1-8 atoms group, the heteroalkyl group contains 1-3 atoms selected from N, O or S; wherein X 3 is a covalent bond or is selected from the following group: hydrogen, C 1 -C 8 alkyl, C 1 -C 8 Alkoxy, C1 - C6cycloalkyl, straight-chain heteroalkyl containing 1-8 atoms, and straight-cyclic-cyclic heteroalkyl containing 1-8 atoms, the heteroalkyl containing selected 1-3 atoms from N, O or S; said C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight containing 1-8 atoms Chain heteroalkyl, and straight-chain-cyclic heteroalkyl containing 1-8 atoms are each independently optionally selected from deuterated, halogen, cyano, nitro, amino, alkyl, carboxy, alkoxy substituted with one or more substituents of a radical or cycloalkyl, the optionally substituted polysarcosine residue comprising
Figure PCTCN2022089726-appb-000033
wherein n2 is a number from 4 to 18, and R is selected from the group consisting of C 1 -C 6 alkyl, C 1 -C 6 cycloalkyl, and C 1 -C 6 alkoxy.
例如，其中，结构单元-X-为任选取代的
Figure PCTCN2022089726-appb-000034
所述任选取代的聚肌氨酸残基包含
Figure PCTCN2022089726-appb-000035
其中n2为4至18的数，R选自以下组：C 1-C 6烷基、C 1-C 6环烷基、和C 1-C 6烷氧基。 For example, wherein the structural unit -X- is optionally substituted
Figure PCTCN2022089726-appb-000034
The optionally substituted polysarcosine residue comprises
Figure PCTCN2022089726-appb-000035
wherein n2 is a number from 4 to 18, and R is selected from the group consisting of C 1 -C 6 alkyl, C 1 -C 6 cycloalkyl, and C 1 -C 6 alkoxy.
例如，其中，结构单元-X-选自以下组：任选取代的
Figure PCTCN2022089726-appb-000036
其中X 1选自以下组：羰基、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；其中X 2选自以下组：氢、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；其中X 3为共价键或选自以下组：氢、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；所述的C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基各自独立地任选地被选自氘代、卤素、氰基、硝基、氨基、烷基、羧基、烷氧基或环烷基的一个或者多个取代基取代，所述任选取代的聚肌氨酸残基包含
Figure PCTCN2022089726-appb-000037
其中n2为4至18的数，R选自以下组：C 1-C 6烷基、C 1-C 6环烷基、和C 1-C 6烷氧基。 For example, wherein the structural unit -X- is selected from the group consisting of optionally substituted
Figure PCTCN2022089726-appb-000036
wherein X 1 is selected from the group consisting of carbonyl, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight chain heteroalkyl containing 1-8 atoms, and Linear-cyclic heteroalkyl groups containing 1-8 atoms containing 1-3 atoms selected from N, O or S ; wherein X is selected from the group consisting of hydrogen, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkane containing 1-8 atoms group, the heteroalkyl group contains 1-3 atoms selected from N, O or S; wherein X 3 is a covalent bond or is selected from the following group: hydrogen, C 1 -C 8 alkyl, C 1 -C 8 Alkoxy, C1 - C6cycloalkyl, straight-chain heteroalkyl containing 1-8 atoms, and straight-cyclic-cyclic heteroalkyl containing 1-8 atoms, the heteroalkyl containing selected 1-3 atoms from N, O or S; said C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight containing 1-8 atoms Chain heteroalkyl, and straight-chain-cyclic heteroalkyl containing 1-8 atoms are each independently optionally selected from deuterated, halogen, cyano, nitro, amino, alkyl, carboxy, alkoxy substituted with one or more substituents of a radical or cycloalkyl, the optionally substituted polysarcosine residue comprising
Figure PCTCN2022089726-appb-000037
wherein n2 is a number from 4 to 18, and R is selected from the group consisting of C 1 -C 6 alkyl, C 1 -C 6 cycloalkyl, and C 1 -C 6 alkoxy.
例如，其中，n2可以为4至18、8至18、4至12或8至12。例如，其中，n2可以为4、5、6、7、8、9、10、11、12、13、14、15、16、17或18。For example, where n2 can be 4 to 18, 8 to 18, 4 to 12, or 8 to 12. For example, where n2 can be 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18.
例如，其中，T可以包含具有抗肿瘤活性的化合物。For example, where T may comprise a compound having antitumor activity.
例如，其中，T可以包含拓扑异构酶抑制剂。For example, where T can comprise a topoisomerase inhibitor.
例如，其中，T可以包含喜树碱类及非喜树碱类拓扑异构酶I抑制剂。For example, where T can include camptothecin-based and non-camptothecin-based topoisomerase I inhibitors.
例如，其中，T可以包含伊沙替康(Exatecan，CAS登录号171335-80-1)和/或贝洛替康(Belotecan，CAS登录号256411-32-2)和/或Genz-644282(CAS登录号529488-28-6)。For example, where T may comprise ixatecan (Exatecan, CAS accession number 171335-80-1) and/or belotecan (Belotecan, CAS accession number 256411-32-2) and/or Genz-644282 (CAS accession number 256411-32-2) Accession No. 529488-28-6).
例如，其中，T可以选自以下组的结构：
Figure PCTCN2022089726-appb-000038
Figure PCTCN2022089726-appb-000039
For example, a structure where T can be selected from the following group:
Figure PCTCN2022089726-appb-000038
Figure PCTCN2022089726-appb-000039
例如，本申请提供一种化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，其中所述化合物可以包含式(C-Trop-2)所示的结构：For example, the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C-Trop-2):
Figure PCTCN2022089726-appb-000040
Figure PCTCN2022089726-appb-000040
其中，Q 1可以包含与巯基偶联后的连接体， Wherein, Q 1 may comprise a linker coupled with a thiol group,
L 1可以包含-L 1a-C(＝O)-，其中，L 1a可以选自以下组：任选取代的C 1-C 7的亚烷基、任选取代的二甘醇基至八甘醇基、任选取代的C 3-C 6亚脂环基、任选取代的亚芳基和任选取代的亚杂芳基； L 1 may comprise -L 1a -C(=O)-, wherein L 1a may be selected from the group consisting of optionally substituted C 1 -C 7 alkylene, optionally substituted diethylene glycol to octaglycol Alcohol, optionally substituted C3 - C6 alicyclic, optionally substituted arylene, and optionally substituted heteroarylene;
L 2可以包含任选取代的选自以下组的氨基酸构成的多肽残基：甘氨酸、苯丙氨酸、缬氨酸、丙氨酸、精氨酸、瓜氨酸、天冬氨酸、天冬酰胺和赖氨酸， L2 may comprise optionally substituted polypeptide residues consisting of amino acids selected from the group consisting of glycine, phenylalanine, valine, alanine, arginine, citrulline, aspartic acid, aspartic acid amide and lysine,
L 3可以包含任选取代的
Figure PCTCN2022089726-appb-000041
L 3 may contain optionally substituted
Figure PCTCN2022089726-appb-000041
其中，L 2可以包含聚肌氨酸残基， wherein L 2 may comprise a polysarcosine residue,
T可以包含拓扑异构酶抑制剂。T may contain a topoisomerase inhibitor.
例如，本申请提供一种化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，其中所述化合物可以包含式(C-Trop-2)所示的结构：For example, the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C-Trop-2):
Figure PCTCN2022089726-appb-000042
Figure PCTCN2022089726-appb-000042
其中，Q 1可以包含与巯基偶联后的连接体， Wherein, Q 1 may comprise a linker coupled with a thiol group,
L 1可以包含-L 1a-C(＝O)-，其中，L 1a可以选自以下组：任选取代的C 1-C 7的亚烷基、任选取代的二甘醇基至八甘醇基、任选取代的C 3-C 6亚脂环基、任选取代的亚芳基和任选取代的亚杂芳基； L 1 may comprise -L 1a -C(=O)-, wherein L 1a may be selected from the group consisting of optionally substituted C 1 -C 7 alkylene, optionally substituted diethylene glycol to octaglycol Alcohol, optionally substituted C3 - C6 alicyclic, optionally substituted arylene, and optionally substituted heteroarylene;
L 2可以包含任选取代的选自以下组的氨基酸构成的多肽残基：甘氨酸、苯丙氨酸、缬氨酸、丙氨酸、精氨酸、瓜氨酸、天冬氨酸、天冬酰胺和赖氨酸， L2 may comprise optionally substituted polypeptide residues consisting of amino acids selected from the group consisting of glycine, phenylalanine, valine, alanine, arginine, citrulline, aspartic acid, aspartic acid amide and lysine,
L 3可以包含任选取代的
Figure PCTCN2022089726-appb-000043
L 3 may contain optionally substituted
Figure PCTCN2022089726-appb-000043
其中，L 3可以包含聚肌氨酸残基， wherein L3 may comprise a polysarcosine residue,
T可以包含拓扑异构酶抑制剂。T may contain a topoisomerase inhibitor.
例如，本申请提供一种化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，其中所述化合物可以包含式(C-Trop-2)所示的结构：For example, the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C-Trop-2):
Figure PCTCN2022089726-appb-000044
Figure PCTCN2022089726-appb-000044
其中，Q 1可以包含任选取代的
Figure PCTCN2022089726-appb-000045
wherein Q 1 may contain optionally substituted
Figure PCTCN2022089726-appb-000045
L 1可以包含-L 1a-C(＝O)-，其中，L 1a可以包含任选取代的C 1-C 7的亚烷基； L 1 may comprise -L 1a -C(=O)-, wherein L 1a may comprise an optionally substituted C 1 -C 7 alkylene group;
L 2可以包含选自以下组的结构：苯丙氨酸-赖氨酸(Phe-Lys)、缬氨酸-丙氨酸(Val-Ala)、缬氨酸-瓜氨酸(Val-Cit)和缬氨酸-赖氨酸(Val-Lys)， L 2 may comprise a structure selected from the group consisting of phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala), valine-citrulline (Val-Cit) and valine-lysine (Val-Lys),
L 3可以包含任选取代的
Figure PCTCN2022089726-appb-000046
L 3 may contain optionally substituted
Figure PCTCN2022089726-appb-000046
其中，L 2可以包含赖氨酸残基，且所述赖氨酸残基可以被任选取代的
Figure PCTCN2022089726-appb-000047
取 代，其中n1为4至18的数， wherein L2 may comprise a lysine residue, and the lysine residue may be optionally substituted
Figure PCTCN2022089726-appb-000047
Substitution, where n1 is a number from 4 to 18,
T可以包含伊沙替康(Exatecan，CAS登录号171335-80-1)和/或贝洛替康(Belotecan，CAS登录号256411-32-2)和/或Genz-644282(CAS登录号529488-28-6)。T may comprise ixatecan (Exatecan, CAS accession number 171335-80-1) and/or belonotecan (Belotecan, CAS accession number 256411-32-2) and/or Genz-644282 (CAS accession number 529488- 28-6).
例如，本申请提供一种化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，其中所述化合物可以包含式(C-Trop-2)所示的结构：For example, the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C-Trop-2):
Figure PCTCN2022089726-appb-000048
Figure PCTCN2022089726-appb-000048
其中，Q 1可以选自以下组：任选取代的
Figure PCTCN2022089726-appb-000049
任选取代的
Figure PCTCN2022089726-appb-000050
任选取代的
Figure PCTCN2022089726-appb-000051
和任选取代的
Figure PCTCN2022089726-appb-000052
wherein Q 1 can be selected from the following group: optionally substituted
Figure PCTCN2022089726-appb-000049
optionally substituted
Figure PCTCN2022089726-appb-000050
optionally substituted
Figure PCTCN2022089726-appb-000051
and optionally substituted
Figure PCTCN2022089726-appb-000052
L 1可以包含-L 1a-C(＝O)-，其中，L 1a可以包含任选取代的C 1-C 7的亚烷基； L 1 may comprise -L 1a -C(=O)-, wherein L 1a may comprise an optionally substituted C 1 -C 7 alkylene group;
L 2可以包含选自以下组的结构：苯丙氨酸-赖氨酸(Phe-Lys)、缬氨酸-丙氨酸(Val-Ala)、缬氨酸-瓜氨酸(Val-Cit)和缬氨酸-赖氨酸(Val-Lys)， L 2 may comprise a structure selected from the group consisting of phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala), valine-citrulline (Val-Cit) and valine-lysine (Val-Lys),
L 3可以包含任选取代的
Figure PCTCN2022089726-appb-000053
L 3 may contain optionally substituted
Figure PCTCN2022089726-appb-000053
其中，L 3可以被任选取代的
Figure PCTCN2022089726-appb-000054
取代，其中n2为4至18的数， wherein L can be optionally substituted
Figure PCTCN2022089726-appb-000054
Substitution, where n2 is a number from 4 to 18,
T可以包含伊沙替康(Exatecan，CAS登录号171335-80-1)和/或贝洛替康(Belotecan，CAS登录号256411-32-2)和/或Genz-644282(CAS登录号529488-28-6)。T may comprise ixatecan (Exatecan, CAS accession number 171335-80-1) and/or belonotecan (Belotecan, CAS accession number 256411-32-2) and/or Genz-644282 (CAS accession number 529488- 28-6).
例如，本申请提供一种化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，其中所述化合物可以包含式(C2-Trop-2)所示的结构：For example, the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C2-Trop-2):
Figure PCTCN2022089726-appb-000055
Figure PCTCN2022089726-appb-000055
其中，L可以含任选取代的亚烷基、任选取代的聚乙二醇基和任选取代的亚脂环基；R 1可以为任选取代的异丙基或任选取代的苄基；R 2可以为任选取代的甲基、任选取代的
Figure PCTCN2022089726-appb-000056
或任选取代的
Figure PCTCN2022089726-appb-000057
R 3可以为氢或任选取代的甲基；其中，R 2可以被任选取代的
Figure PCTCN2022089726-appb-000058
取代，其中n1为4至18的数，T可以包含伊沙替康(Exatecan，CAS登录号171335-80-1)和/或贝洛替康(Belotecan，CAS登录号256411-32-2)和/或Genz-644282(CAS登录号529488-28-6)。 Wherein, L can contain an optionally substituted alkylene group, an optionally substituted polyethylene glycol group and an optionally substituted alicyclic group; R 1 can be an optionally substituted isopropyl group or an optionally substituted benzyl group ; R 2 can be optionally substituted methyl, optionally substituted
Figure PCTCN2022089726-appb-000056
or optionally substituted
Figure PCTCN2022089726-appb-000057
R 3 can be hydrogen or optionally substituted methyl; wherein R 2 can be optionally substituted
Figure PCTCN2022089726-appb-000058
Substitute, where n1 is a number from 4 to 18, and T may comprise ixatecan (Exatecan, CAS Accession No. 171335-80-1) and/or Belotecan (Belotecan, CAS Accession No. 256411-32-2) and /or Genz-644282 (CAS Accession No. 529488-28-6).
例如，本申请提供一种化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，其中所述化合物可以包含式(C2-Trop-2)所示的结构：For example, the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C2-Trop-2):
Figure PCTCN2022089726-appb-000059
Figure PCTCN2022089726-appb-000059
其中，L可以含任选取代的亚烷基、任选取代的聚乙二醇基和任选取代的亚脂环基；R 1可以为任选取代的异丙基或任选取代的苄基；R 2可以为任选取代的甲基、任选取代的
Figure PCTCN2022089726-appb-000060
或任选取代的
Figure PCTCN2022089726-appb-000061
R 3可以为氢或任选取代的甲基；其中，R 2可以被任选取代的
Figure PCTCN2022089726-appb-000062
取代，其中n2为4至18的数，T可以包含伊沙替康(Exatecan，CAS登录号171335-80-1)和/或贝洛替康(Belotecan，CAS登录号256411-32-2)和/或Genz-644282(CAS登录号529488-28-6)。 Wherein, L can contain an optionally substituted alkylene group, an optionally substituted polyethylene glycol group and an optionally substituted alicyclic group; R 1 can be an optionally substituted isopropyl group or an optionally substituted benzyl group ; R 2 can be optionally substituted methyl, optionally substituted
Figure PCTCN2022089726-appb-000060
or optionally substituted
Figure PCTCN2022089726-appb-000061
R 3 can be hydrogen or optionally substituted methyl; wherein R 2 can be optionally substituted
Figure PCTCN2022089726-appb-000062
Substitute, where n2 is a number from 4 to 18, and T may comprise ixatecan (Exatecan, CAS Accession No. 171335-80-1) and/or Belotecan (Belotecan, CAS Accession No. 256411-32-2) and /or Genz-644282 (CAS Accession No. 529488-28-6).
例如，本申请提供一种化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，其中所述化合物可以包含式(C2-Trop-2)所示的结构：For example, the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C2-Trop-2):
Figure PCTCN2022089726-appb-000063
Figure PCTCN2022089726-appb-000063
其中，L可以含任选取代的亚烷基、任选取代的聚乙二醇基和任选取代的亚脂环基；R 1可以为任选取代的异丙基或任选取代的苄基；R 2可以为任选取代的
Figure PCTCN2022089726-appb-000064
R 3可以为氢或任选取代的甲基；其中，R 4可以为任选取代的
Figure PCTCN2022089726-appb-000065
其中n1为4至18的数，T可以包含伊沙替康(Exatecan，CAS登录号171335-80-1)和/或贝洛替康(Belotecan，CAS登录号256411-32-2)和/或Genz-644282(CAS登录号529488-28-6)。 Wherein, L can contain an optionally substituted alkylene group, an optionally substituted polyethylene glycol group and an optionally substituted alicyclic group; R 1 can be an optionally substituted isopropyl group or an optionally substituted benzyl group ; R 2 can be optionally substituted
Figure PCTCN2022089726-appb-000064
R 3 can be hydrogen or optionally substituted methyl; wherein, R 4 can be optionally substituted
Figure PCTCN2022089726-appb-000065
where n1 is a number from 4 to 18, and T may contain ixatecan (Exatecan, CAS accession number 171335-80-1) and/or belotecan (Belotecan, CAS accession number 256411-32-2) and/or Genz-644282 (CAS Accession No. 529488-28-6).
例如，本申请提供一种化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，其中所述化合物可以包含式(C2-Trop-2)所示的结构：For example, the present application provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable form thereof Accepted salts, prodrugs or solvates, wherein the compound may comprise the structure of formula (C2-Trop-2):
Figure PCTCN2022089726-appb-000066
Figure PCTCN2022089726-appb-000066
其中，L可以含任选取代的亚烷基、任选取代的聚乙二醇基和任选取代的亚脂环基；R 1可以为任选取代的异丙基或任选取代的苄基；R 2可以为任选取代的甲基、任选取代的
Figure PCTCN2022089726-appb-000067
或任选取代的
Figure PCTCN2022089726-appb-000068
R 3可以为氢或R 5；其中，R 5可以为任选取代的
Figure PCTCN2022089726-appb-000069
其中n2为4至18的数，T可以包含伊沙替康(Exatecan，CAS登录号171335-80-1)和/或贝洛替康(Belotecan，CAS登录号256411-32-2)和/或Genz-644282(CAS登录号529488-28-6)。 Wherein, L can contain an optionally substituted alkylene group, an optionally substituted polyethylene glycol group and an optionally substituted alicyclic group; R 1 can be an optionally substituted isopropyl group or an optionally substituted benzyl group ; R 2 can be optionally substituted methyl, optionally substituted
Figure PCTCN2022089726-appb-000067
or optionally substituted
Figure PCTCN2022089726-appb-000068
R 3 can be hydrogen or R 5 ; wherein R 5 can be optionally substituted
Figure PCTCN2022089726-appb-000069
where n2 is a number from 4 to 18, and T may comprise ixatecan (Exatecan, CAS accession number 171335-80-1) and/or belotecan (Belotecan, CAS accession number 256411-32-2) and/or Genz-644282 (CAS Accession No. 529488-28-6).
例如，其中所述Ab可以包含抗Trop-2抗体或其抗原结合片段。For example, wherein the Ab can comprise an anti-Trop-2 antibody or antigen-binding fragment thereof.
例如，其中所述抗体可以选自以下组：鼠源抗体、嵌合抗体、人源化抗体和全人源抗 体。For example, wherein the antibody may be selected from the group consisting of murine antibodies, chimeric antibodies, humanized antibodies, and fully human antibodies.
例如，其中所述抗体可以包含单克隆抗体。For example, wherein the antibody may comprise a monoclonal antibody.
例如，其中所述抗体可以包含双特异性抗体。For example, wherein the antibody may comprise a bispecific antibody.
例如，其中所述抗原结合片段可以选自以下组：Fab，Fab′，Fv片段，F(ab') 2，F(ab) 2，scFv，di-scFv，VHH和dAb。 For example, wherein the antigen-binding fragment may be selected from the group consisting of Fab, Fab', Fv fragments, F(ab') 2 , F(ab) 2 , scFv, di-scFv, VHH and dAb.
例如，其中所述Ab的重链HCDR1、HCDR2和HCDR3和轻链LCDR1、LCDR2和LCDR3分别包含抗Trop-2抗体的重链HCDR1、HCDR2和HCDR3和轻链LCDR1、LCDR2和LCDR3。For example, wherein the heavy chains HCDR1, HCDR2 and HCDR3 and light chains LCDR1, LCDR2 and LCDR3 of the Ab comprise heavy chains HCDR1, HCDR2 and HCDR3 and light chains LCDR1, LCDR2 and LCDR3, respectively, of an anti-Trop-2 antibody.
例如，其中所述Ab的重链可变区VH和轻链可变区VL分别包含抗Trop-2抗体的重链可变区VH和轻链可变区VL。For example, wherein the heavy chain variable region VH and light chain variable region VL of the Ab comprise the heavy chain variable region VH and light chain variable region VL, respectively, of an anti-Trop-2 antibody.
例如，其中所述Ab的重链和轻链分别包含抗Trop-2抗体的重链和轻链。For example, wherein the heavy and light chains of the Ab comprise the heavy and light chains, respectively, of an anti-Trop-2 antibody.
例如，其中所述Ab可以包含赛妥珠单抗(Sacituzumab，hRS7)。For example, wherein the Ab can comprise certolizumab (Sacituzumab, hRS7).
例如，其中所述m可以通过选自以下组的方法测定：疏水层析、十二烷基硫酸钠聚丙烯酰胺凝胶电泳和液相质谱。例如，m作为单个单克隆抗体分子与细胞毒性药物偶联后得到的抗体偶联药物中的药物分子与单克隆抗体分子的摩尔比的平均值，m可以为1-8的整数或小数，例如，m可以为约1至约2、约1至约3、约1至约4、约1至约5、约1至约6、约1至约7或约1至约8；例如，m可以为约2至约8、约3至约8、约4至约8、约5至约8、约6至约8、约7至约8，或约1、约2、约3、约4、约5、约6、约7或约8。For example, wherein the m can be determined by a method selected from the group consisting of hydrophobic chromatography, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and liquid phase mass spectrometry. For example, m is the average value of the molar ratio of drug molecules to monoclonal antibody molecules in the antibody-conjugated drug obtained by coupling a single monoclonal antibody molecule with a cytotoxic drug, and m can be an integer or decimal from 1 to 8, such as , m can be about 1 to about 2, about 1 to about 3, about 1 to about 4, about 1 to about 5, about 1 to about 6, about 1 to about 7, or about 1 to about 8; for example, m can be about 2 to about 8, about 3 to about 8, about 4 to about 8, about 5 to about 8, about 6 to about 8, about 7 to about 8, or about 1, about 2, about 3, about 4, About 5, about 6, about 7 or about 8.
另一方面，本申请还提供一种化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，In another aspect, the present application also provides a compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or a mixture thereof A pharmaceutically acceptable salt, prodrug or solvate,
其中，所述化合物包含选自以下组的结构：wherein the compound comprises a structure selected from the group consisting of:
Figure PCTCN2022089726-appb-000070
Figure PCTCN2022089726-appb-000070
Figure PCTCN2022089726-appb-000071
Figure PCTCN2022089726-appb-000071
Figure PCTCN2022089726-appb-000072
Figure PCTCN2022089726-appb-000072
Ab为能够结合Trop-2的配体，m为1-8的数。Ab is a ligand capable of binding Trop-2, and m is a number from 1 to 8.
本申请所述配体可以是蛋白类激素、凝集素、生长因子、抗体或其他能与细胞、受体和/或抗原结合的分子。例如，本申请的配体可以是抗Trop-2抗体或其抗原结合片段。The ligands described herein may be protein hormones, lectins, growth factors, antibodies, or other molecules capable of binding to cells, receptors and/or antigens. For example, the ligand of the present application can be an anti-Trop-2 antibody or an antigen-binding fragment thereof.
在本申请中，所述配体包含抗体轻链可变区VL中的至少一个CDR。本申请所述CDR可以是根据Kabat定义的；也可以是根据Chothia定义的，各种方式定义的CDR序列均包含在本申请的保护范围之内。In the present application, the ligand comprises at least one CDR in the variable region VL of the antibody light chain. The CDRs described in the present application may be defined according to Kabat; or may be defined according to Chothia, and the CDR sequences defined in various ways are all included within the protection scope of the present application.
例如，本申请的抗原结合蛋白可以是包含重链可变区的CDR1-3和轻链可变区的CDR1-3，其中重链可变区的CDR1-3和轻链可变区的CDR1-3可以分别为赛妥珠单抗(Sacituzumab)的重链可变区的CDR1-3和轻链可变区CDR1-3。例如，本申请的抗原结合蛋白可以具有结合TROP2的结合能力。For example, the antigen-binding protein of the present application may comprise CDR1-3 of the heavy chain variable region and CDR1-3 of the light chain variable region, wherein CDR1-3 of the heavy chain variable region and CDR1-3 of the light chain variable region 3 can be CDR1-3 of the heavy chain variable region and CDR1-3 of the light chain variable region of Sacituzumab, respectively. For example, the antigen-binding protein of the present application may have the ability to bind to TROP2.
例如，本申请的抗原结合蛋白可以是包含重链可变区和轻链可变区，其中重链可变区和轻链可变区可以分别为赛妥珠单抗(Sacituzumab)的重链可变区的和轻链可变区。例如，本申请的抗原结合蛋白可以具有结合TROP2的结合能力。For example, the antigen binding protein of the present application can be a heavy chain variable region comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region and the light chain variable region can be the heavy chain variable region of Sacituzumab, respectively variable region and light chain variable region. For example, the antigen-binding protein of the present application may have the ability to bind to TROP2.
例如，本申请的抗原结合蛋白可以是包含重链和轻链，其中重链和轻链可以分别为赛妥珠单抗(Sacituzumab)的重链的和轻链。For example, the antigen binding proteins of the present application may be comprised of heavy and light chains, wherein the heavy and light chains may be the heavy and light chains of Sacituzumab, respectively.
例如，赛妥珠单抗(Sacituzumab)的重链氨基酸序列可以如SEQ ID NO：1所示，赛妥珠单抗(Sacituzumab)的轻链氨基酸序列可以如SEQ ID NO：2所示。For example, the heavy chain amino acid sequence of Sacituzumab can be shown as SEQ ID NO:1, and the light chain amino acid sequence of Sacituzumab can be shown as SEQ ID NO:2.
作为验证本申请化合物显著的优势，例如本申请的配体可以为：例如，赛妥珠单抗(Sacituzumab)的重链氨基酸序列可以如SEQ ID NO：1所示，赛妥珠单抗(Sacituzumab)的轻链氨基酸序列可以如SEQ ID NO：2所示。例如，曲妥珠单抗(Trastuzumab)的重链氨基酸序列可以如SEQ ID NO：3所示，曲妥珠单抗(Trastuzumab)的轻链氨基酸序列可以如SEQ ID NO：4所示。例如，帕妥珠单抗(Pertuzumab)的重链氨基酸序列可以如SEQ ID NO：5所示，帕妥珠单抗(Pertuzumab)的轻链氨基酸序列可以如SEQ ID NO：6所示。例如，恩诺单抗(Enfortumab)的重链氨基酸序列可以如SEQ ID NO：7所示，恩诺单抗(Enfortumab)的轻链氨基酸序列可以如SEQ ID NO：8所示。例如，Patritumab的重链氨基酸序列可以如SEQ ID NO：9所示，Patritumab的轻链氨基酸序列可以如SEQ ID NO：10所示。例如，抗体H01L02的重链氨基酸序列可以如SEQ ID NO：11所示，抗体H01L02的轻链氨基酸序列可以如SEQ ID NO：12所示。As a significant advantage to verify the compounds of the present application, for example, the ligands of the present application can be: for example, the heavy chain amino acid sequence of Sacituzumab can be as shown in SEQ ID NO: 1, ) light chain amino acid sequence can be as shown in SEQ ID NO:2. For example, the heavy chain amino acid sequence of Trastuzumab can be shown in SEQ ID NO:3, and the light chain amino acid sequence of Trastuzumab can be shown in SEQ ID NO:4. For example, the heavy chain amino acid sequence of Pertuzumab can be shown in SEQ ID NO:5, and the light chain amino acid sequence of Pertuzumab can be shown in SEQ ID NO:6. For example, the heavy chain amino acid sequence of Enfortumab can be shown in SEQ ID NO:7, and the light chain amino acid sequence of Enfortumab can be shown in SEQ ID NO:8. For example, the heavy chain amino acid sequence of Patritumab can be set forth in SEQ ID NO:9, and the light chain amino acid sequence of Patritumab can be set forth in SEQ ID NO:10. For example, the heavy chain amino acid sequence of antibody H01L02 can be set forth in SEQ ID NO:11, and the light chain amino acid sequence of antibody H01L02 can be set forth in SEQ ID NO:12.
本申请的抗体可以利用该领域广为周知的技术制备，例如杂交瘤方法、重组DNA技术、噬菌体展示技术、合成技术或该等技术的组合、或该领域己知的其它技术。变体可以是指抗体的氨基酸序列突变体，以及天然多肽的共价衍生物，条件是保留了与天然多肽相当的生物活性。氨基酸序列突变体与天然氨基酸序列的差异一般在于天然氨基酸序列中的一个活多个氨基酸被取代或在多肽序列中缺失和/或插入一个或多个氨基酸。缺失突变体包括天然多肽的片段和N端和/或C端截短突变体。通常氨基酸序列突变体与天然序列相比至少具有70％、75％、80％、85％、90％、95％、98％或99％以上的同源性。Antibodies of the present application can be prepared using techniques well known in the art, such as hybridoma methods, recombinant DNA techniques, phage display techniques, synthetic techniques, or a combination of these techniques, or other techniques known in the art. Variants may refer to amino acid sequence mutants of antibodies, as well as covalent derivatives of the native polypeptide, provided that biological activity comparable to the native polypeptide is retained. Amino acid sequence mutants generally differ from the native amino acid sequence by the substitution of one or more amino acids in the native amino acid sequence or the deletion and/or insertion of one or more amino acids in the polypeptide sequence. Deletion mutants include fragments of the native polypeptide and N-terminal and/or C-terminal truncation mutants. Typically the amino acid sequence mutants have at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% or more homology compared to the native sequence.
由于本申请提供的抗体-药物偶联物可以靶向瞄准特殊的细胞群体，与细胞表面特异蛋白(抗原)结合，从而通过结合物内吞或药物渗入使得药物以活性形式释放到细胞内，因此，本申请的抗体-药物偶联物可以用于治疗目标疾病，本申请的抗体-药物偶联物可以以治疗有效量，通过合适的途径给予受试者(例如人)。需要治疗的受试者可以是有风险，或怀疑患有与特定抗原的活性或表达量有关病症的患者。这样的患者可以通过常规体检来鉴定。Since the antibody-drug conjugates provided in this application can target specific cell populations and bind to specific proteins (antigens) on the cell surface, the drugs can be released into cells in an active form through the endocytosis of the conjugates or drug infiltration. , the antibody-drug conjugate of the present application can be used to treat a target disease, and the antibody-drug conjugate of the present application can be administered to a subject (eg, a human) in a therapeutically effective amount through a suitable route. A subject in need of treatment may be a patient at risk, or suspected of having a disorder associated with the activity or level of expression of a particular antigen. Such patients can be identified by routine physical examination.
当用本申请的抗体-药物偶联物治疗时，可以通过本领域常规的方法进行递送。例如，它可以通过使用脂质体，水凝胶，环糊精，生物可降解的纳米胶囊，或生物粘附性微球被引入到细胞中。或者，所述核酸或载体可在本地通过直接注射或通过使用输注泵递送。其它方 法可以包括通过使用缀合物和生物可降解的聚合物的使用各种运输和载体系统。When treated with the antibody-drug conjugates of the present application, delivery can be performed by methods routine in the art. For example, it can be introduced into cells by using liposomes, hydrogels, cyclodextrins, biodegradable nanocapsules, or bioadhesive microspheres. Alternatively, the nucleic acid or vector can be delivered locally by direct injection or by using an infusion pump. Other methods may include the use of various transport and carrier systems through the use of conjugates and biodegradable polymers.
一方面，本申请提供一种药物组合物，其可以含有本申请中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，以及可以包含任选地药学上可接受的载体。In one aspect, the application provides a pharmaceutical composition, which may contain the compound described in any one of the application, or a tautomer, meso, racemate, enantiomer, The diastereomers, or mixtures thereof, or pharmaceutically acceptable salts, prodrugs or solvates thereof, and may optionally contain a pharmaceutically acceptable carrier.
申请所述的药物组合物除活性化合物外，可以含有一种或多种辅料，所述辅料可以选自以下组的成分：填充剂(稀释剂)、粘合剂、润湿剂、崩解剂和赋形剂等。根据给药方法的不同，组合物可以含有0.1至99重量％的活性化合物。In addition to the active compound, the pharmaceutical composition described in the application may contain one or more excipients, which may be selected from the following group of ingredients: fillers (diluents), binders, wetting agents, disintegrating agents and excipients, etc. Depending on the method of administration, the composition may contain from 0.1 to 99% by weight of active compound.
含活性成分的药物组合物可以是适用于口服的形式，例如片剂、糖锭剂、锭剂、水或油混悬液、可分散粉末或颗粒、乳液、硬或软胶囊，或糖浆剂。可以按照本领域任何已知制备药用组合物的方法制备口服组合物，所述组合物可以含有粘合剂、填充剂、润滑剂、崩解剂或药学上可接受的润湿剂等，所述组合物还可以含有一种或多种选自以下组的成分：甜味剂、矫味剂、着色剂和防腐剂。Pharmaceutical compositions containing the active ingredient may be in a form suitable for oral administration, such as tablets, dragees, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups. Oral compositions may be prepared according to any method known in the art for the preparation of pharmaceutical compositions, which may contain binders, fillers, lubricants, disintegrants or pharmaceutically acceptable wetting agents, etc. The composition may also contain one or more ingredients selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preservatives.
水悬浮液可以含有活性物质和用于混合的适宜制备的水悬浮液的赋形剂。水混悬液也可以含有一种或多种防腐剂，例如一种或多种着色剂、一种或多种矫味剂和一种或多种甜味剂。油混悬液可以通过使活性成分悬浮于植物油中配制而成。油悬浮液可以含有增稠剂。还可以加入上述的甜味剂和矫味剂。Aqueous suspensions may contain the active materials in admixture with excipients suitable for the preparation of aqueous suspensions. The aqueous suspensions may also contain one or more preservatives, such as one or more coloring agents, one or more flavoring agents, and one or more sweetening agents. Oily suspensions can be formulated by suspending the active ingredient in vegetable oils. The oily suspensions may contain thickening agents. The above-mentioned sweetening and flavoring agents may also be added.
药物组合物还可以是用于制备水混悬液的可分散粉末和颗粒提供活性成分，通过加入水混合分散剂、湿润剂、悬浮剂或防腐剂中的一种或多种。也可加入其他赋形剂例如甜味剂、矫味剂和着色剂。通过加入抗氧化剂例如抗坏血酸保存这些组合物。本申请的药物组合物也可以是水包油乳剂的形式。The pharmaceutical compositions may also provide the active ingredient as dispersible powders and granules for preparation of aqueous suspensions by admixing with water one or more of a dispersing agent, wetting agent, suspending agent or preservative. Other excipients such as sweetening, flavouring and colouring agents may also be added. These compositions are preserved by the addition of antioxidants such as ascorbic acid. The pharmaceutical compositions of the present application may also be in the form of oil-in-water emulsions.
药物组合物可以是无菌注射水溶液形式。可以使用的可接受的溶媒或溶剂有水、林格氏液和等渗氯化钠溶液。无菌注射制剂可以是其中活性成分溶于油相的无菌注射水包油微乳。例如将活性成分溶于大豆油和卵磷脂的混合物中。然后可以将油溶液加入水和甘油的混合物中处理形成微乳。可通过局部大量注射，将注射液或微乳注入患者的血流中。或者，可以按可保持本申请化合物恒定循环浓度的方式给予溶液和微乳。为保持这种恒定浓度，可使用连续静脉内递药装置。例如，所述装置可以是静脉注射泵。The pharmaceutical compositions may be in the form of sterile injectable aqueous solutions. Among the acceptable vehicles or solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. The sterile injectable preparation may be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in an oily phase. For example, the active ingredient is dissolved in a mixture of soybean oil and lecithin. The oil solution can then be processed to form a microemulsion by adding it to a mixture of water and glycerol. Injections or microemulsions can be injected into a patient's bloodstream by local bolus injection. Alternatively, solutions and microemulsions can be administered in a manner that maintains a constant circulating concentration of the compounds of the application. To maintain this constant concentration, a continuous intravenous drug delivery device can be used. For example, the device may be an intravenous pump.
药物组合物可以是用于肌内和皮下给药的无菌注射水或油混悬液的形式。可按已知技术，用上述本申请所述适宜的分散剂或湿润剂和悬浮剂配制该混悬液。无菌注射制剂也可以是在肠胃外可接受的无毒稀释剂或溶剂中制备的无菌注射溶液或混悬液。或者，可方便地用无菌固定油作为溶剂或悬浮介质。The pharmaceutical compositions may be in the form of sterile injectable aqueous or oily suspensions for intramuscular and subcutaneous administration. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned herein above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent. Alternatively, sterile fixed oils are conveniently employed as a solvent or suspending medium.
可按用于直肠给药的栓剂形式给予本申请化合物。可通过将药物与在普通温度下为固体但在直肠中为液体，因而在直肠中会溶化而释放药物的适宜的无刺激性赋形剂混合来制备这些药物组合物。此类物质包括可可脂、甘油明胶、氢化植物油、各种分子量的聚乙二醇和聚乙二醇的脂肪酸酯的混合物。The compounds of the present application may be administered in the form of suppositories for rectal administration. These pharmaceutical compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid in the rectum and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, glycerinated gelatin, hydrogenated vegetable oils, polyethylene glycols of various molecular weights and mixtures of fatty acid esters of polyethylene glycols.
如本领域技术人员所熟知的，药物的给药剂量依赖于多种因素，包括但并非限定于以下因素：所用具体化合物的活性、患者的年龄、患者的体重、患者的健康状况、患者的行为、患者的饮食、给药时间、给药方式、排泄的速率、药物的组合等；另外，最佳的治疗方式如治疗的模式、本申请所述化合物或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其可药用的盐，和/或化合物或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其可药用的盐的日用量或可药用的盐的种类可以根据传统的治疗方案来验证。As is well known to those skilled in the art, the dosage of a drug to be administered depends on a variety of factors including, but not limited to, the following factors: the activity of the particular compound used, the age of the patient, the weight of the patient, the health of the patient, the behavior of the patient , the patient's diet, administration time, administration mode, excretion rate, combination of drugs, etc.; in addition, the optimal treatment mode such as the mode of treatment, the compound or its tautomer, meso isomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, and/or compounds or tautomers, mesoisomers, The daily amount of racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, or the species of pharmaceutically acceptable salts can be verified according to conventional therapeutic regimens.
本申请的药物组合物可以含有安全有效量的本申请的抗体-药物偶联物以及药学上可接受的载体。这类载体可以包括(但并不限于)：盐水、缓冲液、葡萄糖、水、甘油、乙醇、及其组合。通常药物制剂应与给药方式相匹配，本申请的药物组合物可以被制成溶液剂形式，例如用生理盐水或含有葡萄糖和其他辅剂的水溶液通过常规方法进行制备。所述的药物组合物可以在无菌条件下制造。活性成分的给药量可以是治疗有效量。The pharmaceutical composition of the present application may contain a safe and effective amount of the antibody-drug conjugate of the present application and a pharmaceutically acceptable carrier. Such carriers can include, but are not limited to, saline, buffers, dextrose, water, glycerol, ethanol, and combinations thereof. Usually the pharmaceutical preparation should match the mode of administration, and the pharmaceutical composition of the present application can be prepared in the form of a solution, for example, prepared by a conventional method with a physiological saline or an aqueous solution containing glucose and other adjuvants. The pharmaceutical composition can be manufactured under sterile conditions. The active ingredient may be administered in a therapeutically effective amount.
本申请所述的抗体-药物偶联物的有效量可以随给药的模式和待治疗的疾病的严重程度等而变化。有效量的选择可以由本领域普通技术人员根据各种因素来确定(例如通过临床试验)。所述的因素可以包括但不限于：所述的双功能抗体偶联物的药代动力学参数例如生物利用率、代谢、半衰期等；患者所要治疗的疾病的严重程度、患者的体重、患者的免疫状况、给药的途径等。通常，当本申请的抗体-药物偶联物每天以合适的剂量给予，可以得到令人满意的效果。例如，由治疗状况的迫切要求，可以每天给予若干次分开的剂量，或将剂量按比例地减少。The effective amount of the antibody-drug conjugates described herein may vary with the mode of administration, the severity of the disease to be treated, and the like. Selection of an effective amount can be determined by one of ordinary skill in the art based on various factors (eg, through clinical trials). The factors may include, but are not limited to: the pharmacokinetic parameters of the diabody conjugate such as bioavailability, metabolism, half-life, etc.; the severity of the disease to be treated by the patient, the weight of the patient, the immune status, route of administration, etc. Generally, when the antibody-drug conjugates of the present application are administered in an appropriate dose per day, satisfactory results can be obtained. For example, several divided doses may be administered daily or the dose may be proportionally reduced as dictated by the exigencies of the therapeutic situation.
本申请化合物可以单独给药，或者可以与其他药学上可接受的治疗剂联合给药。使用药物组合物时，可以是将安全有效量的本申请化合物适用于需要治疗的哺乳动物(如人)，其中施用时剂量可以为药学上认为的有效给药剂量，具体剂量还可以考虑给药途径、病人健康状况等因素。The compounds of the present application may be administered alone, or may be administered in combination with other pharmaceutically acceptable therapeutic agents. When using the pharmaceutical composition, a safe and effective amount of the compound of the present application can be applied to a mammal (such as a human) in need of treatment, and the dose can be a pharmaceutically effective dose when administered, and the specific dose can also be considered. factors such as route, patient's health status, etc.
本申请提供一种本申请的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，和/或本申请的药物组合物在制备可以用于治疗和/或预防肿瘤的药物中的用途。例如，所述肿瘤可以选 自与以下组靶点表达相关的肿瘤：HER2。例如，所述与所述靶点表达相关的肿瘤包含所述靶点高表达的肿瘤和/或所述靶点阳性的肿瘤。例如，所述肿瘤包含实体肿瘤和/或血液肿瘤。例如，所述肿瘤选自以下组：乳腺癌、卵巢癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、急性淋巴细胞性白血病、间变性大细胞淋巴瘤、多发性骨髓瘤、前列腺癌、非小细胞肺癌、小细胞肺癌、恶性黑色素瘤、鳞状细胞癌、胶质母细胞瘤、肾细胞癌、胃肠道肿瘤、胰腺癌、前列腺癌、直结肠、胃癌、神经胶质瘤和间皮瘤。The present application provides a compound of the present application, or a tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable form thereof Use of acceptable salts, prodrugs or solvates, and/or pharmaceutical compositions of the present application in the manufacture of medicaments that can be used to treat and/or prevent tumors. For example, the tumor can be selected from tumors associated with expression of the following group of targets: HER2. For example, the tumor associated with the expression of the target includes a tumor with high expression of the target and/or a tumor positive for the target. For example, the tumor comprises a solid tumor and/or a hematological tumor. For example, the tumor is selected from the group consisting of breast cancer, ovarian cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, acute lymphoblastic leukemia, anaplastic large cell lymphoma, multiple myeloma, prostate cancer, Non-small cell lung cancer, small cell lung cancer, malignant melanoma, squamous cell carcinoma, glioblastoma, renal cell carcinoma, gastrointestinal tumors, pancreatic cancer, prostate cancer, rectal cancer, gastric cancer, glioma and skin tumor.
本申请提供一种本申请的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，和/或本申请的药物组合物，其可以用于治疗和/或预防肿瘤的药物中的用途。例如，所述肿瘤可以选自与以下组靶点表达相关的肿瘤：HER2。例如，所述与所述靶点表达相关的肿瘤包含所述靶点高表达的肿瘤和/或所述靶点阳性的肿瘤。例如，所述肿瘤包含实体肿瘤和/或血液肿瘤。例如，所述肿瘤选自以下组：乳腺癌、卵巢癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、急性淋巴细胞性白血病、间变性大细胞淋巴瘤、多发性骨髓瘤、前列腺癌、非小细胞肺癌、小细胞肺癌、恶性黑色素瘤、鳞状细胞癌、胶质母细胞瘤、肾细胞癌、胃肠道肿瘤、胰腺癌、前列腺癌、直结肠、胃癌、神经胶质瘤和间皮瘤。The present application provides a compound of the present application, or a tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable form thereof Acceptable salts, prodrugs or solvates, and/or pharmaceutical compositions of the present application, which can be used for use in medicaments for the treatment and/or prevention of tumors. For example, the tumor can be selected from tumors associated with expression of the following group of targets: HER2. For example, the tumor associated with the expression of the target includes a tumor with high expression of the target and/or a tumor positive for the target. For example, the tumor comprises a solid tumor and/or a hematological tumor. For example, the tumor is selected from the group consisting of breast cancer, ovarian cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, acute lymphoblastic leukemia, anaplastic large cell lymphoma, multiple myeloma, prostate cancer, Non-small cell lung cancer, small cell lung cancer, malignant melanoma, squamous cell carcinoma, glioblastoma, renal cell carcinoma, gastrointestinal tumors, pancreatic cancer, prostate cancer, rectal cancer, gastric cancer, glioma and skin tumor.
本申请提供一种预防和/或治疗肿瘤的方法，可以包括向受试者施用本申请的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，和/或本申请的药物组合物。例如，所述肿瘤可以选自与以下组靶点表达相关的肿瘤：HER2。例如，所述与所述靶点表达相关的肿瘤包含所述靶点高表达的肿瘤和/或所述靶点阳性的肿瘤。例如，所述肿瘤包含实体肿瘤和/或血液肿瘤。例如，所述肿瘤选自以下组：乳腺癌、卵巢癌、非霍奇金淋巴瘤、霍奇金淋巴瘤、急性淋巴细胞性白血病、间变性大细胞淋巴瘤、多发性骨髓瘤、前列腺癌、非小细胞肺癌、小细胞肺癌、恶性黑色素瘤、鳞状细胞癌、胶质母细胞瘤、肾细胞癌、胃肠道肿瘤、胰腺癌、前列腺癌、直结肠、胃癌、神经胶质瘤和间皮瘤。The present application provides a method of preventing and/or treating tumors, which may include administering to a subject a compound of the present application, or a tautomer, meso, racemate, enantiomer, A diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, and/or the pharmaceutical composition of the present application. For example, the tumor can be selected from tumors associated with expression of the following group of targets: HER2. For example, the tumor associated with the expression of the target includes a tumor with high expression of the target and/or a tumor positive for the target. For example, the tumor comprises a solid tumor and/or a hematological tumor. For example, the tumor is selected from the group consisting of breast cancer, ovarian cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, acute lymphoblastic leukemia, anaplastic large cell lymphoma, multiple myeloma, prostate cancer, Non-small cell lung cancer, small cell lung cancer, malignant melanoma, squamous cell carcinoma, glioblastoma, renal cell carcinoma, gastrointestinal tumors, pancreatic cancer, prostate cancer, rectal cancer, gastric cancer, glioma and skin tumor.
不欲被任何理论所限，下文中的实施例仅仅是为了阐释本申请的化合物、制备方法和用途等，而不用于限制本申请发明的范围。Without intending to be limited by any theory, the following examples are only used to illustrate the compounds, preparation methods, uses, etc. of the present application, and are not intended to limit the scope of the invention of the present application.
实施例Example
实施例1化合物的合成与制备Synthesis and preparation of the compound of Example 1
本申请所述的原料为市售产品，或者通过本领域已知的方法制备或根据本文所述方法制 备。其中，Fmoc是9-芴基甲氧基羰基保护基(9-fluorenylmethyloxycarbonyl)，Boc是叔丁氧基羰基保护基(tert-butoxycarbonyl)，TBDMS/TBS是叔丁基二甲基硅基保护基(tert-butyldimethylsilyl)。The starting materials described herein are commercially available products, either prepared by methods known in the art or prepared according to the methods described herein. Wherein, Fmoc is 9-fluorenylmethoxycarbonyl protecting group (9-fluorenylmethyloxycarbonyl), Boc is tert-butoxycarbonyl protecting group (tert-butoxycarbonyl), TBDMS/TBS is tert-butyldimethylsilyl protecting group ( tert-butyldimethylsilyl).
化合物(A-1)的合成Synthesis of Compound (A-1)
Figure PCTCN2022089726-appb-000073
Figure PCTCN2022089726-appb-000073
合成路线：synthetic route:
Figure PCTCN2022089726-appb-000074
Figure PCTCN2022089726-appb-000074
步骤1：中间体1-1的合成Step 1: Synthesis of Intermediate 1-1
Fmoc-Val-OSu(100g,229mmol)溶解于500毫升四氢呋喃中，分别加入Nε-(叔丁氧基羰基)-L-赖氨酸(59.3g,241mmol)与碳酸氢钠(20.21g,241mmol)在500毫升的水溶液。反应液室温搅拌48小时，检测反应结束。反应液用1N稀盐酸调节pH至6左右，并加入500毫升乙酸乙酯萃取，分离有机相后再经水和饱和盐水各洗一次后，无水硫酸钠干燥，蒸干。剩余物经甲基叔丁基醚重结晶后得产物1-1(107克，收率82％)为白色固体。LC-MS(ESI,m/z)理论值:567.29，实测值:568.26(M+H)。Fmoc-Val-OSu (100g, 229mmol) was dissolved in 500ml of tetrahydrofuran, Nε-(tert-butoxycarbonyl)-L-lysine (59.3g, 241mmol) and sodium bicarbonate (20.21g, 241mmol) were added respectively in 500 ml of aqueous solution. The reaction solution was stirred at room temperature for 48 hours, and the detection reaction was completed. The pH of the reaction solution was adjusted to about 6 with 1N dilute hydrochloric acid, and 500 ml of ethyl acetate was added for extraction. The organic phase was separated, washed once with water and saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness. The residue was recrystallized from methyl tert-butyl ether to give the product 1-1 (107 g, yield 82%) as a white solid. LC-MS (ESI, m/z) theoretical: 567.29, found: 568.26 (M+H).
步骤2：中间体1-2的合成Step 2: Synthesis of Intermediates 1-2
将中间体1-1(60g,106mmol)溶解于二氯甲烷与甲醇(v:v＝2:1，900毫升)的混合溶剂 中。室温下加入对氨基苄醇(19.52g,159mmol)，后再加入EEDQ(39.2g,159mmol)。反应液室温搅拌24小时，反应液减压蒸干，加入乙醚搅拌至固体析出，过滤，固体用乙醚淋洗数次，固体干燥后获得中间体1-2(51克，收率72％)。LC-MS(ESI,m/z)理论值:672.35，实测值:673.37(M+H)。Intermediate 1-1 (60 g, 106 mmol) was dissolved in a mixed solvent of dichloromethane and methanol (v:v=2:1, 900 mL). p-Aminobenzyl alcohol (19.52 g, 159 mmol) was added at room temperature followed by EEDQ (39.2 g, 159 mmol). The reaction solution was stirred at room temperature for 24 hours. The reaction solution was evaporated to dryness under reduced pressure. Diethyl ether was added and stirred until the solid was precipitated. After filtration, the solid was rinsed with diethyl ether for several times, and the solid was dried to obtain Intermediate 1-2 (51 g, yield 72%). LC-MS (ESI, m/z) theoretical: 672.35, found: 673.37 (M+H).
步骤3：中间体1-3的合成Step 3: Synthesis of Intermediates 1-3
将中间体1-2(50g,74.3mmol)溶解于370毫升DMF中。室温下加入二乙胺(78ml,743mmol)。反应液室温搅拌2小时，反应液减压蒸干，加入乙酸乙酸、乙醚搅拌至固体析出，过滤，固体用乙醚淋洗数次，固体干燥后获得中间体1-3(31.5克，收率94％)。LC-MS(ESI,m/z)理论值:450.28，实测值:451.32(M+H)。Intermediate 1-2 (50 g, 74.3 mmol) was dissolved in 370 mL of DMF. Diethylamine (78 ml, 743 mmol) was added at room temperature. The reaction solution was stirred at room temperature for 2 hours, the reaction solution was evaporated to dryness under reduced pressure, acetic acid and diethyl ether were added and stirred until the solid was precipitated, filtered, and the solid was rinsed with diethyl ether several times. %). LC-MS (ESI, m/z) theoretical: 450.28, found: 451.32 (M+H).
步骤4：中间体1-4的合成Step 4: Synthesis of Intermediates 1-4
将中间体1-3(5g,11.10mmol)溶解于100毫升DMF中，室温入加入马来酰亚胺基乙酸琥珀酰亚胺(2.80g,11.10mmol)，反应液室温搅拌过夜，反应液减压蒸干，加入甲基叔丁基醚搅拌至固体析出，过滤，固体用乙醚淋洗数次，固体干燥后获得中间体1-4，直接用于下一步反应。LC-MS(ESI,m/z)理论值:587.30，实测值:588.31(M+H)。Intermediate 1-3 (5 g, 11.10 mmol) was dissolved in 100 ml of DMF, maleimidoacetic acid succinimide (2.80 g, 11.10 mmol) was added at room temperature, the reaction solution was stirred at room temperature overnight, and the reaction solution was reduced. Evaporate to dryness under pressure, add methyl tert-butyl ether and stir until the solid is precipitated, filter, rinse the solid with diethyl ether for several times, and dry the solid to obtain intermediate 1-4, which is directly used in the next reaction. LC-MS (ESI, m/z) theoretical: 587.30, found: 588.31 (M+H).
步骤5：中间体1-5的合成Step 5: Synthesis of Intermediates 1-5
将中间体1-4(2g,3.40mmol)溶解于40毫升DMF中，室温下分别加入DIPEA(1.189ml,6.81mmol)与双(4-硝基苯基)碳酸酯(1.553g,5.10mmol)。氩气下室温搅拌过夜，加入甲基叔丁基醚搅拌至固体析出，过滤，固体用乙醚淋洗数次，固体干燥后获得中间体1-5，直接用于下一步反应。LC-MS(ESI,m/z)理论值:752.30，实测值:753.31(M+H)。Intermediate 1-4 (2g, 3.40mmol) was dissolved in 40ml DMF, DIPEA (1.189ml, 6.81mmol) and bis(4-nitrophenyl)carbonate (1.553g, 5.10mmol) were added at room temperature, respectively. . Stir overnight at room temperature under argon, add methyl tert-butyl ether and stir until the solid precipitates, filter, rinse the solid with ether for several times, and dry the solid to obtain intermediate 1-5, which is directly used in the next reaction. LC-MS (ESI, m/z) theoretical: 752.30, found: 753.31 (M+H).
步骤6：中间体1-6的合成Step 6: Synthesis of Intermediates 1-6
将中间体1-5(142mg,0.188mmol)溶解于400μL无水DMF中，加入100μL无水吡啶，后再加入依沙替康甲磺酸盐(购自上海皓元，100mg,0.188mmol)与HOBt(25.4mg,0.188mmol)。氩气下室温搅拌过夜，反应液经反相HPLC制备纯化得中间体1-6(80mg,收率40％)。LC-MS(ESI,m/z)理论值:1048.43，实测值:1045.45(M+H)。Intermediate 1-5 (142 mg, 0.188 mmol) was dissolved in 400 μL of anhydrous DMF, 100 μL of anhydrous pyridine was added, and then ixatecan mesylate (purchased from Shanghai Haoyuan, 100 mg, 0.188 mmol) and 100 μL of anhydrous pyridine were added. HOBt (25.4 mg, 0.188 mmol). The mixture was stirred at room temperature overnight under argon, and the reaction solution was prepared and purified by reverse-phase HPLC to obtain Intermediate 1-6 (80 mg, yield 40%). LC-MS (ESI, m/z) theoretical: 1048.43, found: 1045.45 (M+H).
步骤7：化合物(A-1)的合成Step 7: Synthesis of Compound (A-1)
将中间体1-6(100mg,0.095mmol)溶解于1mL无水二氯甲烷中，冰浴下加入500μL三氟乙酸后，恢复至室温搅拌30分钟，减压除去溶剂得终产物，后经反相HPLC制备纯化得终产物化合物(A-1)(70mg,收率77％)。LC-MS(ESI,m/z)理论值:1048.43，实测值:1045.45(M+H)。LC-MS(ESI,m/z)理论值:948.38，实测值:949.37(M+H)。Intermediate 1-6 (100 mg, 0.095 mmol) was dissolved in 1 mL of anhydrous dichloromethane, 500 μL of trifluoroacetic acid was added in an ice bath, returned to room temperature and stirred for 30 minutes, and the solvent was removed under reduced pressure to obtain the final product, which was then subjected to reaction The final product compound (A-1) (70 mg, yield 77%) was obtained by HPLC preparation and purification. LC-MS (ESI, m/z) theoretical: 1048.43, found: 1045.45 (M+H). LC-MS (ESI, m/z) theoretical: 948.38, found: 949.37 (M+H).
化合物(A-2)的合成Synthesis of Compound (A-2)
Figure PCTCN2022089726-appb-000075
Figure PCTCN2022089726-appb-000075
化合物(A-2)的合成与化合物(A-1)的合成步骤相同，只是将其中步骤4中的原料马来酰亚胺基乙酸琥珀酰亚胺替换为6-(马来酰亚胺基)己酸琥珀酰亚胺酯，经过数步反应得产物(A-2)为米色无定形粉末。LC-MS(ESI,m/z)理论值:1004.44，实测值:1005.45(M+H)。The synthesis of compound (A-2) is the same as that of compound (A-1), except that the raw material maleimidoacetic acid succinimide in step 4 is replaced with 6-(maleimido group ) caproic acid succinimidyl ester, the product (A-2) is obtained as a beige amorphous powder after several steps of reaction. LC-MS (ESI, m/z) theoretical value: 1004.44, found: 1005.45 (M+H).
化合物(A-3)的合成Synthesis of Compound (A-3)
Figure PCTCN2022089726-appb-000076
Figure PCTCN2022089726-appb-000076
将化合物(A-1)(100mg,0.105mmol)溶解于1毫升无水DMF中，分别加入乙酰化-10聚肌氨酸(Ac-Sar10-COOH，97mg,0.126mmol)，HATU(48mg,0.126mmol)和DIPEA(37μL,0.211mmol)后，室温搅拌过夜，减压除去溶剂得终产物，后经反相HPLC制备纯化得终产物化合物(A-3)(87mg,收率49％)。LC-MS(ESI,m/z)理论值:1700.76，实测值:1701.78(M+H)。Compound (A-1) (100 mg, 0.105 mmol) was dissolved in 1 mL of anhydrous DMF, and acetylated-10 polysarcosine (Ac-Sar10-COOH, 97 mg, 0.126 mmol), HATU (48 mg, 0.126 mmol) were added respectively. mmol) and DIPEA (37 μL, 0.211 mmol), stirred at room temperature overnight, and removed the solvent under reduced pressure to obtain the final product, which was then prepared and purified by reverse-phase HPLC to obtain the final product compound (A-3) (87 mg, yield 49%). LC-MS (ESI, m/z) theoretical: 1700.76, found: 1701.78 (M+H).
化合物(A-4)的合成Synthesis of Compound (A-4)
Figure PCTCN2022089726-appb-000077
Figure PCTCN2022089726-appb-000077
将化合物(A-2)(100mg,0.099mmol)溶解于1毫升无水DMF中，分别加入乙酰化-10聚肌氨酸(Ac-Sar10-COOH，92mg,119mmol)，HATU(45mg,119mmol)和DIPEA(35μL,0.199mmol)后，室温搅拌过夜，减压除去溶剂得终产物，后经反相HPLC制备纯化得终产物化合物(A-4)(95mg,收率54％)。LC-MS(ESI,m/z)理论值:1756.83，实测值:1757.85(M+H)。Compound (A-2) (100 mg, 0.099 mmol) was dissolved in 1 mL of anhydrous DMF, and acetylated-10 polysarcosine (Ac-Sar10-COOH, 92 mg, 119 mmol) and HATU (45 mg, 119 mmol) were added respectively. After mixing with DIPEA (35 μL, 0.199 mmol), stirring at room temperature overnight, the solvent was removed under reduced pressure to obtain the final product, which was then prepared and purified by reverse-phase HPLC to obtain the final product compound (A-4) (95 mg, yield 54%). LC-MS (ESI, m/z) theoretical: 1756.83, found: 1757.85 (M+H).
化合物(A-5)的合成Synthesis of Compound (A-5)
Figure PCTCN2022089726-appb-000078
Figure PCTCN2022089726-appb-000078
化合物(A-5)的合成与化合物(A-4)的合成步骤相同，只是将最后一个步骤中的乙酰化-10聚肌氨酸(Ac-Sar10-COOH)替换为乙酰化-4聚肌氨酸(Ac-Sar4-COOH)，经过数步反应得产物(A-5)为米色无定形粉末。LC-MS(ESI,m/z)理论值:1330.60，实测值:1331.61(M+H)。The synthesis of compound (A-5) is the same as that of compound (A-4), except that the acetylated-10-polysarcosine (Ac-Sar10-COOH) in the last step is replaced by acetylated-4-polysarcosine Amino acid (Ac-Sar4-COOH), the product (A-5) is a beige amorphous powder after several steps of reaction. LC-MS (ESI, m/z) theoretical: 1330.60, found: 1331.61 (M+H).
化合物(A-6)的合成Synthesis of Compound (A-6)
Figure PCTCN2022089726-appb-000079
Figure PCTCN2022089726-appb-000079
化合物(A-6)的合成与化合物(A-4)的合成步骤相同，只是将步骤6中的依沙替康甲磺酸盐替换为贝洛替康盐酸盐(购自上海皓元化学)，经过数步反应得产物(A-6)为白色无定形粉末。LC-MS(ESI,m/z)理论值:1754.87，实测值:1755.88(M+H)。The synthesis of compound (A-6) is the same as the synthesis step of compound (A-4), except that the ixatecan mesylate in step 6 is replaced by belotecan hydrochloride (purchased from Shanghai Haoyuan Chemical). ), the product (A-6) was obtained as a white amorphous powder after several steps of reaction. LC-MS (ESI, m/z) theoretical: 1754.87, found: 1755.88 (M+H).
化合物(A-7)的合成Synthesis of Compound (A-7)
Figure PCTCN2022089726-appb-000080
Figure PCTCN2022089726-appb-000080
化合物(A-7)的合成与化合物(A-4)的合成步骤相同，只是将步骤1中的Fmoc-Val-OSu替换为Fmoc-Phe-OSu，经过数步反应得产物(A-7)为米色无定形粉末。LC-MS(ESI,m/z)理论值:1804.83，实测值:1805.85(M+H)。The synthesis of compound (A-7) is the same as that of compound (A-4), except that Fmoc-Val-OSu in step 1 is replaced by Fmoc-Phe-OSu, and the product (A-7) is obtained after several steps of reaction It is a beige amorphous powder. LC-MS (ESI, m/z) theoretical: 1804.83, found: 1805.85 (M+H).
化合物(A-8)的合成Synthesis of Compound (A-8)
Figure PCTCN2022089726-appb-000081
Figure PCTCN2022089726-appb-000081
化合物(A-8)的合成与化合物(A-7)的合成步骤相同，只是将步骤6中的依沙替康甲磺酸盐替换为贝洛替康盐酸盐(购自上海皓元化学)，经过数步反应得产物(A-8)为白色无定形粉末。LC-MS(ESI,m/z)理论值:1802.87，实测值:1803.86(M+H)。The synthesis of compound (A-8) is the same as the synthesis step of compound (A-7), except that the ixatecan mesylate in step 6 is replaced by belotecan hydrochloride (purchased from Shanghai Haoyuan Chemical). ), the product (A-8) was obtained as a white amorphous powder after several steps of reaction. LC-MS (ESI, m/z) theoretical value: 1802.87, found value: 1803.86 (M+H).
化合物(A-9)的合成Synthesis of Compound (A-9)
Figure PCTCN2022089726-appb-000082
Figure PCTCN2022089726-appb-000082
合成路线：synthetic route:
Figure PCTCN2022089726-appb-000083
Figure PCTCN2022089726-appb-000083
步骤1：中间体9-1的合成Step 1: Synthesis of Intermediate 9-1
将中间体9-1A(1.3g,2.69mmol)与9-1B(1.35g,2.69mmol)溶解于二氯甲烷与甲醇(v:v＝2:1，90毫升)的混合溶剂中。室温下加入对EEDQ(800mg,3.23mmol)。反应液室温搅拌24小时，反应液减压蒸干，经柱层析获得9-1(1.8g，收率69％)。LC-MS(ESI,m/z)理论值:966.49，实测值:967.50(M+H)。Intermediates 9-1A (1.3 g, 2.69 mmol) and 9-1B (1.35 g, 2.69 mmol) were dissolved in a mixed solvent of dichloromethane and methanol (v:v=2:1, 90 mL). Para-EEDQ (800 mg, 3.23 mmol) was added at room temperature. The reaction solution was stirred at room temperature for 24 hours, the reaction solution was evaporated to dryness under reduced pressure, and 9-1 (1.8 g, yield 69%) was obtained by column chromatography. LC-MS (ESI, m/z) theoretical: 966.49, found: 967.50 (M+H).
步骤2：中间体9-2的合成Step 2: Synthesis of Intermediate 9-2
将中间体9-1(900mg,0.93mmol)溶解于20毫升无水四氢呋喃中，氩气氛下冰浴冷却，加入氢氟酸吡啶复合物(1.8g,18.61mmol)，反应液在0度下搅拌2小时，加入水萃灭反应，二氯甲烷萃取，分离出有机层，无水硫酸钠干燥，经柱层析获得中间体9-2(610mg，收率77％)。LC-MS(ESI,m/z)理论值:852.41，实测值:853.43(M+H)。Intermediate 9-1 (900 mg, 0.93 mmol) was dissolved in 20 mL of anhydrous tetrahydrofuran, cooled in an ice bath under argon atmosphere, pyridine hydrofluoric acid complex (1.8 g, 18.61 mmol) was added, and the reaction solution was stirred at 0 degrees After 2 hours, water was added to quench the reaction, extracted with dichloromethane, the organic layer was separated, dried over anhydrous sodium sulfate, and intermediate 9-2 (610 mg, yield 77%) was obtained by column chromatography. LC-MS (ESI, m/z) theoretical: 852.41, found: 853.43 (M+H).
步骤3：中间体9-3的合成Step 3: Synthesis of Intermediate 9-3
将中间体9-2(600mg,0.703mmol)溶解于4毫升无水DMF中，室温下分别加入DIPEA(0.84ml,1.05mmol)与双(4-硝基苯基)碳酸酯(321mg,1.05mmol)。氩气下室温搅拌过夜，减压蒸去溶剂，后加入甲基叔丁基醚搅拌至固体析出，过滤，固体用乙醚淋洗数次，固体干燥后获得中间体9-3，直接用于下一步反应。LC-MS(ESI,m/z)理论值:1017.41，实测值:1018.38(M+H)。Intermediate 9-2 (600 mg, 0.703 mmol) was dissolved in 4 mL of anhydrous DMF, and DIPEA (0.84 mL, 1.05 mmol) and bis(4-nitrophenyl) carbonate (321 mg, 1.05 mmol) were added at room temperature, respectively. ). Stir overnight at room temperature under argon, evaporate the solvent under reduced pressure, then add methyl tert-butyl ether and stir until the solid is precipitated, filter, rinse the solid with diethyl ether several times, and dry the solid to obtain intermediate 9-3, which is directly used in the following one-step reaction. LC-MS (ESI, m/z) theoretical value: 1017.41, found: 1018.38 (M+H).
步骤4：中间体9-4的合成Step 4: Synthesis of Intermediates 9-4
将中间体9-3(300mg,0.295mmol)溶解于400μL无水DMF中，加入100μL无水吡啶，后再加入依沙替康甲磺酸盐(购自上海皓元，157mg,0.295mmol)与HOBt(40mg,0.295mmol)。氩气下室温搅拌过夜，反应液经反相HPLC制备纯化得中间体9-4(160mg,收率41％)。LC-MS(ESI,m/z)理论值:1313.54，实测值:1314.51(M+H)。Intermediate 9-3 (300 mg, 0.295 mmol) was dissolved in 400 μL of anhydrous DMF, 100 μL of anhydrous pyridine was added, and then ixatecan mesylate (purchased from Shanghai Haoyuan, 157 mg, 0.295 mmol) and 100 μL of anhydrous pyridine were added. HOBt (40 mg, 0.295 mmol). The mixture was stirred at room temperature overnight under argon, and the reaction solution was prepared and purified by reverse-phase HPLC to obtain intermediate 9-4 (160 mg, yield 41%). LC-MS (ESI, m/z) theoretical: 1313.54, found: 1314.51 (M+H).
步骤5：中间体9-5的合成Step 5: Synthesis of Intermediates 9-5
将中间体9-4(150mg,0.114mmol)溶解于1毫升DMF中。室温下加入二乙胺(120μL,1.14mmol)。反应液室温搅拌2小时，反应液减压蒸干获得中间体9-5，直接用于下一步反应。LC-MS(ESI,m/z)理论值:1091.48，实测值:1092.51(M+H)。Intermediate 9-4 (150 mg, 0.114 mmol) was dissolved in 1 mL of DMF. Diethylamine (120 μL, 1.14 mmol) was added at room temperature. The reaction solution was stirred at room temperature for 2 hours, and the reaction solution was evaporated to dryness under reduced pressure to obtain intermediate 9-5, which was directly used in the next reaction. LC-MS (ESI, m/z) theoretical value: 1091.48, found value: 1092.51 (M+H).
步骤6：中间体9-6的合成Step 6: Synthesis of Intermediates 9-6
将化合物9-5(120mg,0.110mmol)溶解于1毫升无水DMF中，分别加入乙酰化-10聚肌氨酸(Ac-Sar10-COOH，102mg,0.132mmol)，HATU(50mg,0.132mmol)和DIPEA(38μL,0.22mmol)后，室温搅拌过夜，减压除去溶剂得终产物，后经反相HPLC制备纯化得中间体化合物9-6(105mg,收率52％)。LC-MS(ESI,m/z)理论值:1843.86，实测值:1844.84(M+H)。Compound 9-5 (120 mg, 0.110 mmol) was dissolved in 1 mL of anhydrous DMF, and acetylated-10 polysarcosine (Ac-Sar10-COOH, 102 mg, 0.132 mmol) and HATU (50 mg, 0.132 mmol) were added respectively. After mixing with DIPEA (38 μL, 0.22 mmol), the mixture was stirred at room temperature overnight, and the solvent was removed under reduced pressure to obtain the final product, which was then prepared and purified by reverse-phase HPLC to obtain the intermediate compound 9-6 (105 mg, yield 52%). LC-MS (ESI, m/z) theoretical: 1843.86, found: 1844.84 (M+H).
步骤7：化合物(A-9)的合成Step 7: Synthesis of Compound (A-9)
将中间体9-6(100mg,0.054mmol)溶解于1mL无水二氯甲烷中，冰浴下加入500μL三氟乙酸后，恢复至室温搅拌30分钟，减压除去溶剂得终产物，后经反相HPLC制备纯化得终产物化合物(A-9)(57mg,收率60％)。LC-MS(ESI,m/z)理论值:1048.43，实测值:1045.45(M+H)。LC-MS(ESI,m/z)理论值:1743.81，实测值:1744.85(M+H)。Intermediate 9-6 (100 mg, 0.054 mmol) was dissolved in 1 mL of anhydrous dichloromethane, 500 μL of trifluoroacetic acid was added under an ice bath, returned to room temperature and stirred for 30 minutes, and the solvent was removed under reduced pressure to obtain the final product, which was then subjected to reaction The final product compound (A-9) (57 mg, yield 60%) was obtained by HPLC preparation and purification. LC-MS (ESI, m/z) theoretical: 1048.43, found: 1045.45 (M+H). LC-MS (ESI, m/z) theoretical: 1743.81, found: 1744.85 (M+H).
化合物(A-10)的合成Synthesis of Compound (A-10)
Figure PCTCN2022089726-appb-000084
Figure PCTCN2022089726-appb-000084
化合物(A-10)的合成与化合物(A-9)的合成步骤相同，只是将步骤4中的依沙替康甲磺酸盐替换为贝洛替康盐酸盐(购自上海皓元化学)，经过数步反应得产物(A-10)为白色无定形粉末。LC-MS(ESI,m/z)理论值:1741.85，实测值:1742.83(M+H)。The synthesis of compound (A-10) is the same as that of compound (A-9), except that the ixatecan mesylate in step 4 is replaced by belotecan hydrochloride (purchased from Shanghai Haoyuan Chemical) ), the product (A-10) was obtained as a white amorphous powder after several steps of reaction. LC-MS (ESI, m/z) theoretical: 1741.85, found: 1742.83 (M+H).
化合物(A-11)的合成Synthesis of Compound (A-11)
Figure PCTCN2022089726-appb-000085
Figure PCTCN2022089726-appb-000085
合成路线：synthetic route:
Figure PCTCN2022089726-appb-000086
Figure PCTCN2022089726-appb-000086
步骤1：中间体11-1的合成Step 1: Synthesis of Intermediate 11-1
将中间体11-1A(Mc-Val-Ala-OH，购自上海皓元化学，2.4g,6.29mmol)与11-1B(3.18g,6.29mmol)溶解于二氯甲烷与甲醇(v:v＝2:1，90毫升)的混合溶剂中。室温下加入对EEDQ(1.86g,7.55mmol)。反应液室温搅拌24小时，反应液减压蒸干，经柱层析获得11-1(3.9g，收率71％)。LC-MS(ESI,m/z)理论值:867.46，实测值:868.49(M+H)。Intermediates 11-1A (Mc-Val-Ala-OH, purchased from Shanghai Haoyuan Chemical, 2.4 g, 6.29 mmol) and 11-1B (3.18 g, 6.29 mmol) were dissolved in dichloromethane and methanol (v:v = 2:1, 90 ml) in a mixed solvent. Para-EEDQ (1.86 g, 7.55 mmol) was added at room temperature. The reaction solution was stirred at room temperature for 24 hours, the reaction solution was evaporated to dryness under reduced pressure, and 11-1 (3.9 g, yield 71%) was obtained by column chromatography. LC-MS (ESI, m/z) theoretical: 867.46, found: 868.49 (M+H).
步骤2：中间体11-2的合成Step 2: Synthesis of Intermediate 11-2
将中间体11-1(2g,2.3mmol)溶解于50毫升无水四氢呋喃中，氩气氛下冰浴冷却，加入氢氟酸吡啶复合物(4.6g,46mmol)，反应液在0度下搅拌2小时，加入水萃灭反应，二氯甲烷萃取，分离出有机层，无水硫酸钠干燥，经柱层析获得中间体11-2(1.1g，收率76％)。LC-MS(ESI,m/z)理论值:629.34，实测值:630.31(M+H)。Intermediate 11-1 (2 g, 2.3 mmol) was dissolved in 50 mL of anhydrous tetrahydrofuran, cooled in an ice bath under argon atmosphere, pyridine hydrofluoric acid complex (4.6 g, 46 mmol) was added, and the reaction solution was stirred at 0 degrees for 2 After 1 hour, water was added to quench the reaction, extracted with dichloromethane, the organic layer was separated, dried over anhydrous sodium sulfate, and intermediate 11-2 (1.1 g, yield 76%) was obtained by column chromatography. LC-MS (ESI, m/z) theoretical: 629.34, found: 630.31 (M+H).
步骤3：中间体11-3的合成Step 3: Synthesis of Intermediate 11-3
将中间体11-2(700mg,1.11mmol)溶解于4毫升无水DMF中，室温下分别加入DIPEA(0.39ml,2.23mmol)与双(4-硝基苯基)碳酸酯(406mg,1.33mmol)。氩气下室温搅拌过夜，减压蒸去溶剂，后加入甲基叔丁基醚搅拌至固体析出，过滤，固体用乙醚淋洗数次，固体干燥后获得中间体11-3，直接用于下一步反应。LC-MS(ESI,m/z)理论值:794.35，实测值:795.41(M+H)。Intermediate 11-2 (700 mg, 1.11 mmol) was dissolved in 4 mL of anhydrous DMF, and DIPEA (0.39 mL, 2.23 mmol) and bis(4-nitrophenyl) carbonate (406 mg, 1.33 mmol) were added at room temperature, respectively. ). Stir overnight at room temperature under argon, evaporate the solvent under reduced pressure, then add methyl tert-butyl ether and stir until the solid is precipitated, filter, rinse the solid with ether several times, and dry the solid to obtain intermediate 11-3, which is directly used in the following one-step reaction. LC-MS (ESI, m/z) theoretical: 794.35, found: 795.41 (M+H).
步骤4：中间体11-4的合成Step 4: Synthesis of Intermediate 11-4
将中间体11-3(300mg,0.44mmol)溶解于400μL无水DMF中，加入100μL无水吡啶，后再加入依沙替康甲磺酸盐(购自上海皓元，234mg,0.44mmol)与HOBt(60mg,0.44mmol)。氩气下室温搅拌过夜，反应液经反相HPLC制备纯化得中间体11-4(230mg,收率48％)。LC-MS(ESI,m/z)理论值:1090.48，实测值:1091.53(M+H)。Intermediate 11-3 (300 mg, 0.44 mmol) was dissolved in 400 μL of anhydrous DMF, 100 μL of anhydrous pyridine was added, and then ixatecan mesylate (purchased from Shanghai Haoyuan, 234 mg, 0.44 mmol) and 100 μL of anhydrous pyridine were added. HOBt (60 mg, 0.44 mmol). The mixture was stirred overnight at room temperature under argon, and the reaction solution was prepared and purified by reverse-phase HPLC to obtain Intermediate 11-4 (230 mg, yield 48%). LC-MS (ESI, m/z) theoretical: 1090.48, found: 1091.53 (M+H).
步骤5：中间体11-5的合成Step 5: Synthesis of Intermediate 11-5
将中间体11-4(200mg,0.183mmol)溶解于1mL无水二氯甲烷中，冰浴下加入300μL三氟乙酸后，恢复至室温搅拌30分钟，减压除去溶剂得终产物得中间体11-5的三氟乙酸盐，未经进一步纯化直接用于下一步反应。LC-MS(ESI,m/z)理论值:990.43，实测值:991.47(M+H)。Intermediate 11-4 (200 mg, 0.183 mmol) was dissolved in 1 mL of anhydrous dichloromethane, 300 μL of trifluoroacetic acid was added under ice bath, returned to room temperature and stirred for 30 minutes, and the solvent was removed under reduced pressure to obtain the final product to obtain intermediate 11 The trifluoroacetate salt of -5 was used in the next reaction without further purification. LC-MS (ESI, m/z) theoretical: 990.43, found: 991.47 (M+H).
步骤6：化合物(A-11)的合成Step 6: Synthesis of Compound (A-11)
将化合物11-5(120mg,0.109mmol)溶解于1毫升无水DMF中，分别加入乙酰化-10聚肌氨酸(Ac-Sar10-COOH，84mg,0.109mmol)，HATU(50mg,0.130mmol)和DIPEA(38μL,0.22mmol)后，室温搅拌过夜，减压除去溶剂得终产物，后经反相HPLC制备纯化得化合物(A-11)(74mg,收率38％)。LC-MS(ESI,m/z)理论值:1742.81，实测值:1743.85(M+H)。Compound 11-5 (120 mg, 0.109 mmol) was dissolved in 1 mL of anhydrous DMF, and acetylated-10 polysarcosine (Ac-Sar10-COOH, 84 mg, 0.109 mmol) and HATU (50 mg, 0.130 mmol) were added respectively. After mixing with DIPEA (38 μL, 0.22 mmol), the mixture was stirred at room temperature overnight, and the solvent was removed under reduced pressure to obtain the final product, which was then prepared and purified by reverse-phase HPLC to obtain compound (A-11) (74 mg, yield 38%). LC-MS (ESI, m/z) theoretical: 1742.81, found: 1743.85 (M+H).
化合物(A-12)的合成Synthesis of Compound (A-12)
Figure PCTCN2022089726-appb-000087
Figure PCTCN2022089726-appb-000087
化合物(A-12)的合成与化合物(A-11)的合成步骤相同，只是将步骤6中的原料化合物Ac-Sar10-COOH替换为Ac-Sar4-COOH，经过数步反应得产物(A-12)为米色无定形粉末。LC-MS(ESI,m/z)理论值:1316.59，实测值:1317.62(M+H)。The synthesis of compound (A-12) is the same as that of compound (A-11), except that the raw material compound Ac-Sar10-COOH in step 6 is replaced by Ac-Sar4-COOH, and the product (A- 12) is a beige amorphous powder. LC-MS (ESI, m/z) theoretical: 1316.59, found: 1317.62 (M+H).
化合物(A-13)的合成Synthesis of Compound (A-13)
Figure PCTCN2022089726-appb-000088
Figure PCTCN2022089726-appb-000088
化合物(A-13)的合成与化合物(A-11)的合成步骤相同，只是将步骤1中的原料化合物11-1A替换为Mc-Val-Cit-OH(购自上海皓元化学)，经过数步反应得产物(A-13)为米色无定形粉末。LC-MS(ESI,m/z)理论值:1828.86，实测值:1829.88(M+H)。The synthesis of compound (A-13) is the same as that of compound (A-11), except that the raw material compound 11-1A in step 1 is replaced by Mc-Val-Cit-OH (purchased from Shanghai Haoyuan Chemical), after The product (A-13) was obtained as a beige amorphous powder after several steps of reaction. LC-MS (ESI, m/z) theoretical value: 1828.86, found value: 1829.88 (M+H).
化合物(A-14)的合成Synthesis of Compound (A-14)
Figure PCTCN2022089726-appb-000089
Figure PCTCN2022089726-appb-000089
化合物(A-14)的合成与化合物(A-11)的合成步骤相同，只是将步骤1中的原料化合物11-1A(Mc-VA-OH)替换为Mc-GGFG-OH(购自上海皓元化学)，经过数步反应得产物(A- 13)为米色无定形粉末。LC-MS(ESI,m/z)理论值:1890.84，实测值:1891.90(M+H)。The synthesis of compound (A-14) is the same as that of compound (A-11), except that the raw material compound 11-1A (Mc-VA-OH) in step 1 is replaced by Mc-GGFG-OH (purchased from Shanghai Hao) Yuan Chemistry), after several steps of reaction, the product (A-13) is a beige amorphous powder. LC-MS (ESI, m/z) theoretical: 1890.84, found: 1891.90 (M+H).
化合物(A-15)的合成Synthesis of Compound (A-15)
Figure PCTCN2022089726-appb-000090
Figure PCTCN2022089726-appb-000090
合成路线：synthetic route:
Figure PCTCN2022089726-appb-000091
Figure PCTCN2022089726-appb-000091
步骤1：中间体15-2的合成Step 1: Synthesis of Intermediate 15-2
将中间体15-1A(2.3g,8.57mmol)与15-1B(3.74g,8.57mmol)溶解于二氯甲烷与甲醇(v:v＝2:1，90毫升)的混合溶剂中。室温下加入对EEDQ(4.24g,17.15mmol)。反应液室温搅拌24小时，反应液减压蒸干，经柱层析获得中间体15-2(3.23g,4.70mmol,54.9％yield)。LC-MS(ESI,m/z)理论值:686.27，实测值:687.21(M+H)。Intermediates 15-1A (2.3 g, 8.57 mmol) and 15-1B (3.74 g, 8.57 mmol) were dissolved in a mixed solvent of dichloromethane and methanol (v:v=2:1, 90 mL). Para-EEDQ (4.24 g, 17.15 mmol) was added at room temperature. The reaction solution was stirred at room temperature for 24 hours, the reaction solution was evaporated to dryness under reduced pressure, and intermediate 15-2 (3.23 g, 4.70 mmol, 54.9% yield) was obtained by column chromatography. LC-MS (ESI, m/z) theoretical: 686.27, found: 687.21 (M+H).
步骤2：中间体15-3的合成Step 2: Synthesis of Intermediate 15-3
将中间体15-2(2.3g,3.35mmol)溶解于40毫升无水DMF中，室温下分别加入DIPEA(1.170ml,6.70mmol)与双(4-硝基苯基)碳酸酯(1.528g,5.02mmol)。氩气下室温搅拌过夜，减 压蒸去溶剂，后加入甲基叔丁基醚搅拌至固体析出，过滤，固体用乙醚淋洗数次，固体干燥后获得中间体15-3(2.1g,2.465mmol,收率73.6％)，直接用于下一步反应。Intermediate 15-2 (2.3g, 3.35mmol) was dissolved in 40ml of anhydrous DMF, DIPEA (1.170ml, 6.70mmol) and bis(4-nitrophenyl)carbonate (1.528g, 5.02 mmol). Stir at room temperature overnight under argon, evaporate the solvent under reduced pressure, then add methyl tert-butyl ether and stir until the solid is precipitated, filter, rinse the solid with diethyl ether several times, and dry the solid to obtain Intermediate 15-3 (2.1 g, 2.465 g) mmol, yield 73.6%), which was directly used in the next reaction.
步骤3：中间体15-4的合成Step 3: Synthesis of Intermediate 15-4
将中间体15-3(400mg,0.470mmol)溶解于4mL无水DMF中，加入1mL无水吡啶，后再加入依沙替康游离碱(购自上海皓元化学)(204mg,0.470mmol)与HOBt(63.4mg,0.470mmol)。氩气下室温搅拌过夜，反应液蒸干后，柱层析纯化(DCM:MeOH＝30:1)得中间体15-4(244mg,0.213mmol,收率45.3％)。LC-MS(ESI,m/z)理论值:1147.41，实测值:1148.48(M+H)。The intermediate 15-3 (400 mg, 0.470 mmol) was dissolved in 4 mL of anhydrous DMF, 1 mL of anhydrous pyridine was added, and then ixatecan free base (purchased from Shanghai Haoyuan Chemical) (204 mg, 0.470 mmol) and 1 mL of anhydrous pyridine were added. HOBt (63.4 mg, 0.470 mmol). The mixture was stirred at room temperature overnight under argon, and the reaction solution was evaporated to dryness and purified by column chromatography (DCM:MeOH=30:1) to obtain intermediate 15-4 (244 mg, 0.213 mmol, yield 45.3%). LC-MS (ESI, m/z) theoretical: 1147.41, found: 1148.48 (M+H).
步骤4：中间体15-5的合成Step 4: Synthesis of Intermediate 15-5
将中间体15-4(200mg,0.183mmol)溶解于1mL无水二氯甲烷中，冰浴下加入300μL三氟乙酸后，恢复至室温搅拌30分钟，减压除去溶剂得终产物得中间体15-5的三氟乙酸盐，未经进一步纯化直接用于下一步反应。LC-MS(ESI,m/z)理论值:1047.36，实测值:1048.40(M+H)。Intermediate 15-4 (200 mg, 0.183 mmol) was dissolved in 1 mL of anhydrous dichloromethane, 300 μL of trifluoroacetic acid was added under ice bath, returned to room temperature and stirred for 30 minutes, and the solvent was removed under reduced pressure to obtain the final product to obtain intermediate 15 The trifluoroacetate salt of -5 was used in the next reaction without further purification. LC-MS (ESI, m/z) theoretical: 1047.36, found: 1048.40 (M+H).
步骤5：化合物A-15的合成Step 5: Synthesis of Compound A-15
将中间体化合物15-5(150mg,0.131mmol)溶解于1毫升无水DMF中，分别加入乙酰化-10聚肌氨酸(Ac-Sar10-COOH)(101mg,0.131mmol)，DIPEA(114μl,0.654mmol)和HATU(74.6mg,0.196mmol)后，室温搅拌过夜，减压除去溶剂得终产物，后经反相HPLC制备纯化得化合物A-15(107mg,0.059mmol,收率45.4％)。LC-MS(ESI,m/z)理论值:1799.74，实测值:1800.77(M+H)。The intermediate compound 15-5 (150 mg, 0.131 mmol) was dissolved in 1 ml of anhydrous DMF, and acetylated-10 polysarcosine (Ac-Sar10-COOH) (101 mg, 0.131 mmol), DIPEA (114 μl, 0.654 mmol) and HATU (74.6 mg, 0.196 mmol), stirred at room temperature overnight, the solvent was removed under reduced pressure to obtain the final product, which was then prepared and purified by reverse-phase HPLC to obtain compound A-15 (107 mg, 0.059 mmol, yield 45.4%). LC-MS (ESI, m/z) theoretical: 1799.74, found: 1800.77 (M+H).
化合物(A-16)的合成Synthesis of Compound (A-16)
Figure PCTCN2022089726-appb-000092
Figure PCTCN2022089726-appb-000092
化合物(A-16)的合成与化合物(A-15)的合成步骤相同，只是将步骤3中的依沙替康甲磺酸盐替换为贝洛替康盐酸盐(购自上海皓元化学)，经过数步反应得产物(A-16)为米色无定形粉末。LC-MS(ESI,m/z)理论值:1797.78，实测值:1798.81(M+H)。The synthesis of compound (A-16) is the same as that of compound (A-15), except that the ixatecan mesylate in step 3 is replaced by belotecan hydrochloride (purchased from Shanghai Haoyuan Chemical). ), the product (A-16) was obtained as a beige amorphous powder after several steps of reaction. LC-MS (ESI, m/z) theoretical: 1797.78, found: 1798.81 (M+H).
化合物(A-17)的合成Synthesis of Compound (A-17)
Figure PCTCN2022089726-appb-000093
Figure PCTCN2022089726-appb-000093
化合物(A-17)的合成与化合物(A-11)的合成步骤相同，只是将步骤4中的依沙替康甲磺酸盐替换为Genz-644282(购自上海皓元化学)，经过数步反应得产物(A-17)为米色无定形粉末。LC-MS(ESI,m/z)理论值:1714.80，实测值:1715.83(M+H)。The synthesis of compound (A-17) is the same as that of compound (A-11), except that the ixatecan mesylate in step 4 is replaced by Genz-644282 (purchased from Shanghai Haoyuan Chemical), after several The product (A-17) obtained from the first reaction was a beige amorphous powder. LC-MS (ESI, m/z) theoretical: 1714.80, found: 1715.83 (M+H).
化合物(A-18)的合成Synthesis of Compound (A-18)
Figure PCTCN2022089726-appb-000094
Figure PCTCN2022089726-appb-000094
合成路线：synthetic route:
Figure PCTCN2022089726-appb-000095
Figure PCTCN2022089726-appb-000095
步骤1：中间体18-2的合成Step 1: Synthesis of Intermediate 18-2
将中间体18-1A(购自上海毕得医药，4.94g,14.82mmol)与15-1B(依据WO2016038383中的方法合成，6.04g,14.82mmol)溶解于二氯甲烷与甲醇(v:v＝2:1，120毫升)的混合溶剂中。室温下加入对EEDQ(7.33g,29.6mmol)。反应液室温搅拌24小时，反应液减压蒸干，产物经乙醚搅拌过滤后真空干燥得中间体18-2(6.3g,59％yield)。LC-MS(ESI,m/z)理论值:726.40， 实测值:727.41(M+H)。Intermediates 18-1A (purchased from Shanghai Bide Medicine, 4.94 g, 14.82 mmol) and 15-1B (synthesized according to the method in WO2016038383, 6.04 g, 14.82 mmol) were dissolved in dichloromethane and methanol (v:v= 2:1, 120 mL) in a mixed solvent. Para-EEDQ (7.33 g, 29.6 mmol) was added at room temperature. The reaction solution was stirred at room temperature for 24 hours, the reaction solution was evaporated to dryness under reduced pressure, the product was stirred and filtered with ether, and then vacuum-dried to obtain Intermediate 18-2 (6.3 g, 59% yield). LC-MS (ESI, m/z) theoretical: 726.40, found: 727.41 (M+H).
步骤2：中间体18-3的合成Step 2: Synthesis of Intermediate 18-3
将中间体18-2(3.0g,4.13mmol)溶解于50毫升THF中，加入10％Pd-C(800mg)，常压氢化8小时后，TLC检测反应完全，过滤除去钯碳，反应液蒸干后直接用于下一步反应。LC-MS(ESI,m/z)理论值:636.36，实测值:637.34(M+H)。Intermediate 18-2 (3.0 g, 4.13 mmol) was dissolved in 50 mL of THF, 10% Pd-C (800 mg) was added, and after 8 hours of hydrogenation at atmospheric pressure, TLC detected that the reaction was complete, the palladium carbon was removed by filtration, and the reaction solution was evaporated. After drying, it was directly used in the next reaction. LC-MS (ESI, m/z) theoretical: 636.36, found: 637.34 (M+H).
步骤3：中间体18-4的合成Step 3: Synthesis of Intermediate 18-4
将中间体化合物18-3(2.6g,4.08mmol)溶解于100毫升无水DMF中，分别加入N-(2-氨基乙基)马来酰亚胺盐酸盐(721mg,4.08mmol)，DIPEA(1.42mL,8.17mmol)和HATU(2.0g,5.31mmol)后，室温搅拌过夜，减压除去溶剂得终产物，后经柱层析纯化得化合物18-4(2.2g,71％yield)。LC-MS(ESI,m/z)理论值:758.40，实测值:759.42(M+H)。The intermediate compound 18-3 (2.6 g, 4.08 mmol) was dissolved in 100 mL of anhydrous DMF, and N-(2-aminoethyl)maleimide hydrochloride (721 mg, 4.08 mmol), DIPEA was added respectively. (1.42 mL, 8.17 mmol) and HATU (2.0 g, 5.31 mmol), stirred at room temperature overnight, and removed the solvent under reduced pressure to obtain the final product, which was purified by column chromatography to obtain compound 18-4 (2.2 g, 71% yield). LC-MS (ESI, m/z) theoretical: 758.40, found: 759.42 (M+H).
步骤4：中间体18-5的合成Step 4: Synthesis of Intermediate 18-5
将中间体18-4(1.0g,1.32mmol)溶解于20毫升无水四氢呋喃中，氩气氛下冰浴冷却，加入氢氟酸吡啶复合物(2.6g,26.4mmol)，反应液在0度下搅拌2小时，加入水萃灭反应，二氯甲烷萃取，分离出有机层，无水硫酸钠干燥，经柱层析获得中间体18-5(570mg，收率67％)。LC-MS(ESI,m/z)理论值:644.32，实测值:645.35(M+H)。Intermediate 18-4 (1.0 g, 1.32 mmol) was dissolved in 20 mL of anhydrous tetrahydrofuran, cooled in an ice bath under argon atmosphere, and pyridine hydrofluoric acid complex (2.6 g, 26.4 mmol) was added, and the reaction solution was at 0 degrees. Stir for 2 hours, add water to quench the reaction, extract with dichloromethane, separate the organic layer, dry over anhydrous sodium sulfate, and obtain Intermediate 18-5 (570 mg, yield 67%) by column chromatography. LC-MS (ESI, m/z) theoretical: 644.32, found: 645.35 (M+H).
步骤5：中间体18-6的合成Step 5: Synthesis of Intermediate 18-6
将中间体18-5(500mg,0.776mmol)溶解于10毫升无水DMF中，室温下分别加入DIPEA(271μl,1.55mmol)与双(4-硝基苯基)碳酸酯(236mg0.776mmol)。氩气下室温搅拌过夜，减压蒸去溶剂，后加入甲基叔丁基醚搅拌至固体析出，过滤，固体用乙醚淋洗数次，固体干燥后获得中间体18-6(530mg,84％yield)，直接用于下一步反应。Intermediate 18-5 (500 mg, 0.776 mmol) was dissolved in 10 mL of dry DMF, and DIPEA (271 μl, 1.55 mmol) and bis(4-nitrophenyl) carbonate (236 mg, 0.776 mmol) were added at room temperature, respectively. Stir at room temperature overnight under argon, evaporate the solvent under reduced pressure, then add methyl tert-butyl ether and stir until the solid is precipitated, filter, rinse the solid with diethyl ether several times, and dry the solid to obtain Intermediate 18-6 (530 mg, 84% yield), which is directly used in the next reaction.
步骤6：中间体18-7的合成Step 6: Synthesis of Intermediate 18-7
将中间体18-6(400mg,0.494mmol)溶解于4mL无水DMF中，加入1mL无水吡啶，后再加入依沙替康甲磺酸盐(购自上海皓元，263mg,0.494mmol)与HOBt(66.7mg,0.494mmol)。氩气下室温搅拌过夜，反应液经反相HPLC制备纯化得中间体18-7(320mg,收率58％)。LC-MS(ESI,m/z)理论值:1105.46，实测值:1106.48(M+H)。Intermediate 18-6 (400 mg, 0.494 mmol) was dissolved in 4 mL of anhydrous DMF, 1 mL of anhydrous pyridine was added, and then ixatecan mesylate (purchased from Shanghai Haoyuan, 263 mg, 0.494 mmol) and 1 mL of anhydrous pyridine were added. HOBt (66.7 mg, 0.494 mmol). The mixture was stirred at room temperature overnight under argon, and the reaction solution was prepared and purified by reverse-phase HPLC to obtain intermediate 18-7 (320 mg, yield 58%). LC-MS (ESI, m/z) theoretical: 1105.46, found: 1106.48 (M+H).
步骤7：中间体18-8的合成Step 7: Synthesis of Intermediate 18-8
将中间体15-4(300mg,0.27mmol)溶解于3mL无水二氯甲烷中，冰浴下加入1mL三氟乙酸后，恢复至室温搅拌30分钟，减压除去溶剂得终产物得中间体18-8的三氟乙酸盐，未经进一步纯化直接用于下一步反应。LC-MS(ESI,m/z)理论值:1005.40，实测值:1006.41(M+H)。Intermediate 15-4 (300 mg, 0.27 mmol) was dissolved in 3 mL of anhydrous dichloromethane, 1 mL of trifluoroacetic acid was added under ice bath, returned to room temperature and stirred for 30 minutes, and the solvent was removed under reduced pressure to obtain the final product to obtain intermediate 18 The trifluoroacetate salt of -8 was used in the next reaction without further purification. LC-MS (ESI, m/z) theoretical value: 1005.40, found value: 1006.41 (M+H).
步骤8：化合物A-18的合成Step 8: Synthesis of Compound A-18
将中间体化合物15-5(300mg,0.268mmol)溶解于5毫升无水DMF中，分别加入乙酰化-10聚肌氨酸(Ac-Sar10-COOH)(206mg,0.268mmol)，DIPEA(94μl,0.536mmol)和HATU(122mg,0.321mmol)后，室温搅拌过夜，减压除去溶剂得终产物，后经反相HPLC制备纯化得化合物A-18(170mg,36％yield)。LC-MS(ESI,m/z)理论值:1757.78，实测值:1758.82(M+H)。The intermediate compound 15-5 (300 mg, 0.268 mmol) was dissolved in 5 ml of anhydrous DMF, and acetylated-10 polysarcosine (Ac-Sar10-COOH) (206 mg, 0.268 mmol), DIPEA (94 μl, 0.536 mmol) and HATU (122 mg, 0.321 mmol), stirred overnight at room temperature, and removed the solvent under reduced pressure to obtain the final product, which was then prepared and purified by reverse-phase HPLC to obtain compound A-18 (170 mg, 36% yield). LC-MS (ESI, m/z) theoretical: 1757.78, found: 1758.82 (M+H).
化合物(A-19)的合成Synthesis of Compound (A-19)
Figure PCTCN2022089726-appb-000096
Figure PCTCN2022089726-appb-000096
化合物(A-19)的合成与化合物(A-18)的合成步骤相同，只是将步骤3中的N-(2-氨基乙基)马来酰亚胺盐酸盐替换为2-(methylsulfonyl)benzo[d]thiazol-6-amine，经过数步反应得产物(A-19)。LC-MS(ESI,m/z)理论值:1845.73，实测值:1846.75(M+H)。The synthesis of compound (A-19) is the same as that of compound (A-18), except that N-(2-aminoethyl)maleimide hydrochloride in step 3 is replaced by 2-(methylsulfonyl) benzo[d]thiazol-6-amine, the product (A-19) is obtained after several steps of reaction. LC-MS (ESI, m/z) theoretical: 1845.73, found: 1846.75 (M+H).
化合物(A-20)的合成Synthesis of Compound (A-20)
Figure PCTCN2022089726-appb-000097
Figure PCTCN2022089726-appb-000097
化合物(A-20)的合成与化合物(A-18)的合成步骤相同，只是将步骤3中的N-(2-氨基乙基)马来酰亚胺盐酸盐替换为4-(5-(methylsulfonyl)-1,3,4-oxadiazol-2-yl)aniline，经过数步反应得产物(A-20)。LC-MS(ESI,m/z)理论值:1856.76，实测值:1857.80(M+H)。The synthesis of compound (A-20) is the same as that of compound (A-18), except that N-(2-aminoethyl)maleimide hydrochloride in step 3 is replaced by 4-(5- (methylsulfonyl)-1,3,4-oxadiazol-2-yl)aniline, the product (A-20) was obtained after several steps of reaction. LC-MS (ESI, m/z) theoretical: 1856.76, found: 1857.80 (M+H).
化合物(A-21)的合成Synthesis of Compound (A-21)
Figure PCTCN2022089726-appb-000098
Figure PCTCN2022089726-appb-000098
化合物(A-21)的合成与化合物(A-11)的合成步骤相同，只是将步骤1中的中间体
Figure PCTCN2022089726-appb-000099
替换为
Figure PCTCN2022089726-appb-000100
经过数步反应得产物(A-21)。LC-MS(ESI,m/z)理论值:1742.81，实测值:1743.85(M+H)。 The synthesis of compound (A-21) is the same as that of compound (A-11), except that the intermediate in step 1
Figure PCTCN2022089726-appb-000099
replace with
Figure PCTCN2022089726-appb-000100
The product (A-21) is obtained after several steps of reaction. LC-MS (ESI, m/z) theoretical: 1742.81, found: 1743.85 (M+H).
化合物(A-22)的合成Synthesis of Compound (A-22)
Figure PCTCN2022089726-appb-000101
Figure PCTCN2022089726-appb-000101
化合物(A-22)的合成与化合物(A-21)的合成步骤相同，只是将步骤1中的原料化合物Mc-Val-Ala-OH替换为Mc-Val-Cit-OH，经过数步反应得产物(A-22)为米色无定形粉末。LC-MS(ESI,m/z)理论值:1828.86，实测值:1829.87(M+H)。The synthesis of compound (A-22) is the same as that of compound (A-21), except that the raw material compound Mc-Val-Ala-OH in step 1 is replaced by Mc-Val-Cit-OH, and after several steps of reaction, the obtained The product (A-22) was a beige amorphous powder. LC-MS (ESI, m/z) theoretical value: 1828.86, found value: 1829.87 (M+H).
化合物(A-23)的合成Synthesis of Compound (A-23)
Figure PCTCN2022089726-appb-000102
Figure PCTCN2022089726-appb-000102
化合物(A-23)的合成与化合物(A-21)的合成步骤相同，只是将步骤1中的原料化合物Mc-Val-Ala-OH替换为Mc-GGFG-OH，经过数步反应得产物(A-23)为米色无定形粉末。LC-MS(ESI,m/z)理论值:1890.84，实测值:1891.86(M+H)。The synthesis of compound (A-23) is the same as that of compound (A-21), except that the raw material compound Mc-Val-Ala-OH in step 1 is replaced by Mc-GGFG-OH, and the product ( A-23) is a beige amorphous powder. LC-MS (ESI, m/z) theoretical value: 1890.84, found value: 1891.86 (M+H).
化合物(A-28)的合成Synthesis of Compound (A-28)
Figure PCTCN2022089726-appb-000103
Figure PCTCN2022089726-appb-000103
合成路线：synthetic route:
Figure PCTCN2022089726-appb-000104
Figure PCTCN2022089726-appb-000104
步骤1：中间体28-1的合成Step 1: Synthesis of Intermediate 28-1
称取6-nitroisobenzofuran-1(3H)-one(10g,55.8mmol)与tert-butyl(2-aminoethyl)carbamate(9.84g,61.4mmol)在100毫升圆底烧瓶中，加热至90度搅拌过夜。加入甲基叔丁基醚(MTBE)搅拌，过滤，固体用MTBE洗涤多次后，真空干燥获得中间体28-1(13.6g，收率72％)。LC-MS(ESI,m/z)理论值:339.14，实测值:340.17(M+H)。Weigh 6-nitroisobenzofuran-1(3H)-one (10 g, 55.8 mmol) and tert-butyl(2-aminoethyl)carbamate (9.84 g, 61.4 mmol) in a 100-mL round-bottom flask, heat to 90 degrees and stir overnight. Methyl tert-butyl ether (MTBE) was added, stirred, filtered, and the solid was washed with MTBE for several times, and then dried under vacuum to obtain intermediate 28-1 (13.6 g, yield 72%). LC-MS (ESI, m/z) theoretical: 339.14, found: 340.17 (M+H).
步骤2：中间体28-2的合成Step 2: Synthesis of Intermediate 28-2
将中间体28-1(10g,29.5mmol)与咪唑(8.02g,118mmol)溶解于300毫升二氯甲烷中，冰浴冷却下加入TBS-Cl(6.66g,44.2mmol)，室温下搅拌过夜。反应液加入水淬灭反应，分离有机相后再经水和饱和盐水各洗一次后，无水硫酸钠干燥，蒸干。粗品柱层析纯化得中间体28-2(12.3g，收率92％)。LC-MS(ESI,m/z)理论值:453.23，实测值:454.33(M+H)。Intermediate 28-1 (10 g, 29.5 mmol) and imidazole (8.02 g, 118 mmol) were dissolved in 300 mL of dichloromethane, TBS-Cl (6.66 g, 44.2 mmol) was added under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was added with water to quench the reaction, the organic phase was separated, washed once with water and saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness. The crude product was purified by column chromatography to obtain intermediate 28-2 (12.3 g, yield 92%). LC-MS (ESI, m/z) theoretical: 453.23, found: 454.33 (M+H).
步骤3：中间体28-3的合成Step 3: Synthesis of Intermediate 28-3
将中间体28-2(6.0g,13.23mmol)溶解于THF:EtOH(1:1)(500mL)，氩气下加入1克10％Pd-C，再加入甲酸铵(8.34g,132mmol)。室温搅拌过夜，过滤除去钯碳，蒸干滤液，加入二氯甲烷与水，分离有机相后再经水和饱和盐水各洗一次后，无水硫酸钠干燥，蒸干，得中间体28-3(5.6g,100％)。LC-MS(ESI,m/z)理论值:423.26，实测值:424.31(M+H)。Intermediate 28-2 (6.0 g, 13.23 mmol) was dissolved in THF:EtOH (1:1) (500 mL) and 1 g of 10% Pd-C was added under argon followed by ammonium formate (8.34 g, 132 mmol). Stir overnight at room temperature, remove palladium carbon by filtration, evaporate the filtrate to dryness, add dichloromethane and water, separate the organic phase, wash once with water and saturated brine, dry over anhydrous sodium sulfate, and evaporate to dryness to obtain Intermediate 28-3 (5.6 g, 100%). LC-MS (ESI, m/z) theoretical: 423.26, found: 424.31 (M+H).
步骤4：中间体28-4的合成Step 4: Synthesis of Intermediate 28-4
将中间体28-3(5.5g,12.98mmol)与Fmoc-Val-Alal-OH(5.33g,12.98mmol)溶解于二氯甲烷与甲醇(v:v＝2:1，300毫升)的混合溶剂中。室温下加入EEDQ(4.82g,19.47mmol)。反应液室温搅拌24小时，反应液减压蒸干，加入乙醚搅拌至固体析出，过滤，固体用乙醚淋洗数次，固体干燥后获得中间体28-4(6.4克，收率60％)。LC-MS(ESI,m/z)理论值:815.43，实测值:816.48(M+H)。Intermediate 28-3 (5.5 g, 12.98 mmol) and Fmoc-Val-Alal-OH (5.33 g, 12.98 mmol) were dissolved in a mixed solvent of dichloromethane and methanol (v:v=2:1, 300 mL) middle. EEDQ (4.82 g, 19.47 mmol) was added at room temperature. The reaction solution was stirred at room temperature for 24 hours. The reaction solution was evaporated to dryness under reduced pressure. Diethyl ether was added and stirred until the solid was precipitated. After filtration, the solid was rinsed with diethyl ether for several times, and the solid was dried to obtain Intermediate 28-4 (6.4 g, yield 60%). LC-MS (ESI, m/z) theoretical: 815.43, found: 816.48 (M+H).
步骤5：中间体28-5的合成Step 5: Synthesis of Intermediate 28-5
将中间体28-4(6.0g,7.35mmol)溶解于100毫升DMF中。室温下加入二乙胺(7.68ml,73.5mmol)。反应液室温搅拌2小时，反应液减压蒸干，加入乙酸乙酸、乙醚搅拌至固体析出，过滤，固体用乙醚淋洗数次，固体干燥后获得中间体28-5(4.3克，收率98％)。LC-MS(ESI,m/z)理论值:593.36，实测值:594.40(M+H)。Intermediate 28-4 (6.0 g, 7.35 mmol) was dissolved in 100 mL of DMF. Diethylamine (7.68 ml, 73.5 mmol) was added at room temperature. The reaction solution was stirred at room temperature for 2 hours, the reaction solution was evaporated to dryness under reduced pressure, acetic acid and diethyl ether were added and stirred until the solid was precipitated, filtered, and the solid was rinsed with diethyl ether several times. %). LC-MS (ESI, m/z) theoretical: 593.36, found: 594.40 (M+H).
步骤6：中间体28-6的合成Step 6: Synthesis of Intermediate 28-6
将中间体28-5(4.0g,6.74mmol)溶解于100毫升DMF中，室温入加入马来酰亚胺基乙酸琥珀酰亚胺(2.07g,6.74mmol)，反应液室温搅拌过夜，反应液减压蒸干，加入甲基叔丁基醚搅拌至固体析出，过滤，固体用乙醚淋洗数次，固体干燥后获得中间体28-6，直接用于下一步反应。LC-MS(ESI,m/z)理论值:786.43，实测值:787.45(M+H)。Intermediate 28-5 (4.0 g, 6.74 mmol) was dissolved in 100 ml of DMF, maleimidoacetic acid succinimide (2.07 g, 6.74 mmol) was added at room temperature, and the reaction solution was stirred at room temperature overnight. Evaporate to dryness under reduced pressure, add methyl tert-butyl ether and stir until the solid precipitates, filter, rinse the solid with ether for several times, and dry the solid to obtain intermediate 28-6, which is directly used in the next reaction. LC-MS (ESI, m/z) theoretical: 786.43, found: 787.45 (M+H).
步骤7：中间体28-7的合成Step 7: Synthesis of Intermediate 28-7
将中间体28-6(3.5g,4.45mmol)溶解于100毫升无水四氢呋喃中，氩气氛下冰浴冷却，加入氢氟酸吡啶复合物(8.8g,89mmol)，反应液在0度下搅拌2小时，加入水萃灭反应，二氯甲烷萃取，分离出有机层，无水硫酸钠干燥，经柱层析获得中间体28-7(1.6g，收率53％)。LC-MS(ESI,m/z)理论值:672.35，实测值:673.37(M+H)。Intermediate 28-6 (3.5 g, 4.45 mmol) was dissolved in 100 mL of anhydrous tetrahydrofuran, cooled in an ice bath under argon atmosphere, pyridine hydrofluoric acid complex (8.8 g, 89 mmol) was added, and the reaction solution was stirred at 0 degrees After 2 hours, water was added to quench the reaction, extracted with dichloromethane, the organic layer was separated, dried over anhydrous sodium sulfate, and intermediate 28-7 (1.6 g, yield 53%) was obtained by column chromatography. LC-MS (ESI, m/z) theoretical: 672.35, found: 673.37 (M+H).
步骤8：中间体28-8的合成Step 8: Synthesis of Intermediate 28-8
将中间体28-7(1.0g,1.49mmol)溶解于20毫升无水DMF中，室温下分别加入DIPEA(0.519ml,2.97mmol)与双(4-硝基苯基)碳酸酯(678mg,2.23mmol)。氩气下室温搅拌过夜，减压蒸去溶剂，后加入甲基叔丁基醚搅拌至固体析出，过滤，固体用乙醚淋洗数次，固体干燥后获得中间体28-8，直接用于下一步反应。LC-MS(ESI,m/z)理论值:837.35，实测值:838.36(M+H)。Intermediate 28-7 (1.0 g, 1.49 mmol) was dissolved in 20 mL of anhydrous DMF, DIPEA (0.519 mL, 2.97 mmol) and bis(4-nitrophenyl) carbonate (678 mg, 2.23 mmol) were added at room temperature, respectively. mmol). Stir overnight at room temperature under argon, evaporate the solvent under reduced pressure, then add methyl tert-butyl ether and stir until the solid is precipitated, filter, rinse the solid with ether for several times, and dry the solid to obtain intermediate 28-8, which is directly used in the following one-step reaction. LC-MS (ESI, m/z) theoretical: 837.35, found: 838.36 (M+H).
步骤9：中间体28-9的合成Step 9: Synthesis of Intermediate 28-9
将中间体28-8(300mg,0.358mmol)溶解于4mL无水DMF中，加入1mL无水吡啶，后再加入依沙替康甲磺酸盐(购自上海皓元，190mg,0.358mmol)与HOBt(55mg,0.358mmol)。氩气下室温搅拌过夜，反应液经反相HPLC制备纯化得中间体28-9(234mg,收率58％)。LC-MS(ESI,m/z)理论值:1133.49，实测值:1134.52(M+H)。Intermediate 28-8 (300 mg, 0.358 mmol) was dissolved in 4 mL of anhydrous DMF, 1 mL of anhydrous pyridine was added, and then ixatecan mesylate (purchased from Shanghai Haoyuan, 190 mg, 0.358 mmol) and 1 mL of anhydrous pyridine were added. HOBt (55 mg, 0.358 mmol). The mixture was stirred at room temperature overnight under argon, and the reaction solution was prepared and purified by reverse-phase HPLC to obtain intermediate 28-9 (234 mg, yield 58%). LC-MS (ESI, m/z) theoretical: 1133.49, found: 1134.52 (M+H).
步骤10：中间体28-10的合成Step 10: Synthesis of Intermediate 28-10
将中间体28-10(200mg,0.176mmol)溶解于1mL无水二氯甲烷中，冰浴下加入300μL三氟乙酸后，恢复至室温搅拌30分钟，减压除去溶剂得终产物得中间体28-10的三氟乙酸盐，未经进一步纯化直接用于下一步反应。LC-MS(ESI,m/z)理论值:1033.43，实测值:1034.45(M+H)。Intermediate 28-10 (200 mg, 0.176 mmol) was dissolved in 1 mL of anhydrous dichloromethane, 300 μL of trifluoroacetic acid was added under ice bath, returned to room temperature and stirred for 30 minutes, and the solvent was removed under reduced pressure to obtain the final product to obtain intermediate 28 The trifluoroacetate salt of -10 was used in the next reaction without further purification. LC-MS (ESI, m/z) theoretical: 1033.43, found: 1034.45 (M+H).
步骤11：化合物A28的合成Step 11: Synthesis of Compound A28
将步骤10中所获得的化合物28-10溶解于1毫升无水DMF中，分别加入乙酰化-10聚肌氨酸(Ac-Sar10-COOH，136mg,0.176mmol)，HATU(87mg,0.229mmol)和DIPEA(154μL,0.88mmol)后，室温搅拌过夜，减压除去溶剂得终产物，后经反相HPLC制备纯化得化合物(A-11)(135mg,收率43％)。LC-MS(ESI,m/z)理论值:1785.82，实测值:1786.85(M+H)。Compound 28-10 obtained in step 10 was dissolved in 1 mL of anhydrous DMF, and acetylated-10 polysarcosine (Ac-Sar10-COOH, 136 mg, 0.176 mmol) and HATU (87 mg, 0.229 mmol) were added respectively. After mixing with DIPEA (154 μL, 0.88 mmol), the mixture was stirred at room temperature overnight, and the solvent was removed under reduced pressure to obtain the final product, which was then prepared and purified by reverse-phase HPLC to obtain compound (A-11) (135 mg, yield 43%). LC-MS (ESI, m/z) theoretical: 1785.82, found: 1786.85 (M+H).
化合物(A-29)的合成Synthesis of Compound (A-29)
Figure PCTCN2022089726-appb-000105
Figure PCTCN2022089726-appb-000105
化合物(A-29)的合成与化合物(A-11)的合成步骤相同，只是将步骤4中的依沙替康 甲磺酸盐替换为贝洛替康盐酸盐(购自上海皓元化学)，经过数步反应得产物(A-29)为白色无定形粉末。LC-MS(ESI,m/z)理论值:1740.85，实测值:1741.82(M+H)。The synthesis of compound (A-29) is the same as the synthesis step of compound (A-11), except that the ixatecan mesylate in step 4 is replaced by belotecan hydrochloride (purchased from Shanghai Haoyuan Chemical). ), the product (A-29) was obtained as a white amorphous powder after several steps of reaction. LC-MS (ESI, m/z) theoretical: 1740.85, found: 1741.82 (M+H).
A-11分子对应的PEG衍生物AP-1的合成Synthesis of PEG Derivative AP-1 Corresponding to A-11 Molecule
Figure PCTCN2022089726-appb-000106
Figure PCTCN2022089726-appb-000106
化合物(AP-1)的合成与化合物(A-11)的合成步骤相同，只是将步骤1中的原料化合物Ac-Sar10-COOH替换为m-PEG8-acid(CAS 1093647-41-6)，经过数步反应得产物(AP-1)为米色无定形粉末。LC-MS(ESI,m/z)理论值:1384.64，实测值:1385.66(M+H)。The synthesis of compound (AP-1) is the same as that of compound (A-11), except that the raw material compound Ac-Sar10-COOH in step 1 is replaced by m-PEG8-acid (CAS 1093647-41-6), after The product (AP-1) was obtained as a beige amorphous powder after several steps of reaction. LC-MS (ESI, m/z) theoretical: 1384.64, found: 1385.66 (M+H).
对照品MC-Val-Cit-PABC-DX8951的合成Synthesis of reference substance MC-Val-Cit-PABC-DX8951
参照文献WO2020233174A1方法，Mc-Val-Cit-OH(购自上海皓元化学)与双(4-硝基苯基)碳酸酯反应，获得活化硝基酯后并进一步与依沙替康甲磺酸盐(购自上海皓元)反应获得对照品化合物MC-Val-Cit-PAB-DX8951。Referring to the method of WO2020233174A1, Mc-Val-Cit-OH (purchased from Shanghai Haoyuan Chemical) reacts with bis(4-nitrophenyl) carbonate to obtain activated nitroester and further reacts with ixatecan methanesulfonic acid Salt (purchased from Shanghai Haoyuan) was reacted to obtain the reference compound MC-Val-Cit-PAB-DX8951.
实施例2ADC制备的通用方法Example 2 General method for ADC preparation
将抗体或抗原结合片段，例如将靶向HER2的Trastuzumab原液(重链序列如SEQ ID NO：3所示，轻链序列如SEQ ID NO：4所示)和/或例如靶向Trop-2的hRS7单抗(Sacituzumab)，用50mM磷酸二氢钾-氢氧化钠(KH 2PO4-NaOH)/150mM氯化钠(NaCl)/1mM二乙基三胺五乙酸(DTPA)，pH 7反应缓冲液稀释至2mg/mL，加入6.0倍过量摩尔比的三(2-羧乙基)膦盐酸盐(TCEP)，反应液于35℃搅动2.5小时。 The antibody or antigen-binding fragment, eg, a stock solution of Trastuzumab targeting HER2 (heavy chain sequence is shown in SEQ ID NO: 3, light chain sequence is shown in SEQ ID NO: 4) and/or, for example, targeting Trop-2. hRS7 mAb (Sacituzumab) in 50 mM potassium dihydrogen phosphate-sodium hydroxide (KH 2 PO4-NaOH)/150 mM sodium chloride (NaCl)/1 mM diethyltriaminepentaacetic acid (DTPA), pH 7 reaction buffer Dilute to 2 mg/mL, add tris(2-carboxyethyl)phosphine hydrochloride (TCEP) in an excess molar ratio of 6.0 times, and stir the reaction solution at 35° C. for 2.5 hours.
将上述反应液冷至8℃，未经纯化加入适量的二甲基乙酰胺(DMA)，再分别加入6-15倍过量摩尔比的对照品药物分子或药物连接子缀合物A1～A29(10mg/ml预先溶在DMA中)，保证反应体系中DMA的体积占比不超过20％，于37℃搅动3小时进行偶联。The above reaction solution was cooled to 8°C, an appropriate amount of dimethylacetamide (DMA) was added without purification, and then 6-15 times excess molar ratio of reference substance drug molecules or drug linker conjugates A1 to A29 ( 10 mg/ml pre-dissolved in DMA) to ensure that the volume ratio of DMA in the reaction system does not exceed 20%, and the coupling was performed by stirring at 37° C. for 3 hours.
采用脱盐柱将偶联反应混合物用pH 6.0的组氨酸-醋酸/蔗糖凝胶过滤纯化，根据UV280紫外吸收值收集出峰样品。然后经由0.15微米孔径的过滤装置除菌，-60℃保存。The coupling reaction mixture was purified by gel filtration of pH 6.0 histidine-acetic acid/sucrose using a desalting column, and the peak samples were collected according to the UV280 absorption value. It was then sterilized through a 0.15 micron pore size filter and stored at -60°C.
抗体偶联物1(对照ADC1)的制备Preparation of Antibody Conjugate 1 (Control ADC1)
参照上述通用方法将10倍过量的对照品MC-Val-Cit-PAB-DX8951与还原后的Trastuzumab单抗偶联获得对应的抗体偶联物1，通过尺寸排阻层析(SEC-HPLC)分析聚合体的含量，分析图谱见图1，聚体1的比例约为26％，聚体2的比例约为20％。通过UV分析获得载药量(DAR)约为4～5。Referring to the above general method, a 10-fold excess of the reference substance MC-Val-Cit-PAB-DX8951 was coupled with the reduced Trastuzumab monoclonal antibody to obtain the corresponding antibody conjugate 1, which was analyzed by size exclusion chromatography (SEC-HPLC) The content of the polymer, the analytical map is shown in Figure 1, the proportion of the polymer 1 is about 26%, and the proportion of the polymer 2 is about 20%. The drug loading (DAR) was about 4-5 obtained by UV analysis.
抗体偶联物2(对照ADC2)的制备Preparation of Antibody Conjugate 2 (Control ADC2)
参照上述通用方法将12倍过量的对照品Deruxtecan(购自上海皓元化学)与还原后的Trastuzumab单抗偶联获得对应的抗体偶联物2，通过尺寸排阻层析(SEC-HPLC)分析聚合体的含量，分析图谱见图2，聚体的比例约为2％。通过疏水作用高效液相色谱(HIC-HPLC)分析获得载药量(DAR)约为7-8，分析图谱见图3。Referring to the above general method, a 12-fold excess of the reference substance Deruxtecan (purchased from Shanghai Haoyuan Chemical) was coupled with the reduced Trastuzumab monoclonal antibody to obtain the corresponding antibody conjugate 2, which was analyzed by size exclusion chromatography (SEC-HPLC) The content of the polymer, the analytical map is shown in Figure 2, and the proportion of the polymer is about 2%. The drug loading (DAR) was about 7-8 by hydrophobic interaction high performance liquid chromatography (HIC-HPLC) analysis, and the analytical spectrum is shown in Figure 3.
抗体偶联物3的制备Preparation of Antibody Conjugate 3
参照上述通用方法将8倍过量化合物(A-2)与还原后的Trastuzumab单抗偶联获得对应的抗体偶联物3，通过尺寸排阻层析(SEC-HPLC)分析聚合体的含量，分析图谱见图4。聚体1的比例约为6.8％，聚体2的比例约为7.7％。通过UV分析获得载药量(DAR)约为6～7。Referring to the above general method, an 8-fold excess of compound (A-2) was coupled with the reduced Trastuzumab monoclonal antibody to obtain the corresponding antibody conjugate 3, and the content of the polymer was analyzed by size exclusion chromatography (SEC-HPLC). The map is shown in Figure 4. The proportion of aggregate 1 is about 6.8%, and the proportion of aggregate 2 is about 7.7%. The drug loading (DAR) was about 6-7 obtained by UV analysis.
抗体偶联物4的制备Preparation of Antibody Conjugate 4
参照上述通用方法将将12倍过量化合物(A-4)与还原后的Trastuzumab单抗偶联获得载药量(DAR)约为7～8的抗体偶联物4，通过尺寸排阻层析(SEC-HPLC)分析聚合体的含量，分析图谱见图5。聚体1的比例约为0.3％。通过疏水作用高效液相色谱(HIC-HPLC)分析获得载药量(DAR)约为7-8，分析图谱见图6。Referring to the above general method, a 12-fold excess of compound (A-4) was conjugated with the reduced Trastuzumab monoclonal antibody to obtain antibody conjugate 4 with a drug loading (DAR) of about 7 to 8. Size exclusion chromatography ( SEC-HPLC) to analyze the content of the polymer, and the analytical spectrum is shown in Figure 5. The proportion of aggregate 1 is about 0.3%. The drug loading (DAR) was about 7-8 by hydrophobic interaction high performance liquid chromatography (HIC-HPLC) analysis, and the analytical spectrum is shown in Figure 6.
抗体偶联物5的制备Preparation of Antibody Conjugate 5
参照上述通用方法将12倍过量化合物(A-11)与还原后的Trastuzumab单抗偶联获得对应的抗体偶联物5，通过尺寸排阻层析(SEC-HPLC)分析聚合体的含量，分析图谱见图7，抗体偶联物5单体的纯度为99.41％。通过疏水作用高效液相色谱(HIC-HPLC)分析获得载药量(DAR)约为7-8，分析图谱见图8。Referring to the above general method, a 12-fold excess of compound (A-11) was coupled with the reduced Trastuzumab monoclonal antibody to obtain the corresponding antibody conjugate 5, and the content of the polymer was analyzed by size exclusion chromatography (SEC-HPLC). The map is shown in Figure 7, and the purity of the antibody conjugate 5 monomer is 99.41%. The drug loading (DAR) was about 7-8 obtained by hydrophobic interaction high performance liquid chromatography (HIC-HPLC) analysis, and the analytical spectrum is shown in Figure 8 .
抗体偶联物6的制备Preparation of Antibody Conjugate 6
参照上述通用方法将12倍过量化合物(A-14)与还原后的Trastuzumab单抗偶联获得对应的抗体偶联物6，通过尺寸排阻层析(SEC-HPLC)分析聚合体的含量，分析图谱见图9，抗体偶联物6单体的纯度为99.60％。通过疏水作用高效液相色谱(HIC-HPLC)分析获得载药量(DAR)约为7-8，分析图谱见图10。Referring to the above general method, 12-fold excess compound (A-14) was coupled with the reduced Trastuzumab monoclonal antibody to obtain the corresponding antibody conjugate 6, and the content of the polymer was analyzed by size exclusion chromatography (SEC-HPLC). The map is shown in Figure 9, and the purity of the antibody conjugate 6 monomer is 99.60%. The drug loading (DAR) was obtained by hydrophobic interaction high performance liquid chromatography (HIC-HPLC) to be about 7-8, and the analytical spectrum is shown in Figure 10.
抗体偶联物7的制备Preparation of Antibody Conjugate 7
参照上述通用方法将15倍过量化合物(A-24)与还原后的Trastuzumab单抗偶联获得对应的抗体偶联物7，通过尺寸排阻层析(SEC-HPLC)分析聚合体的含量，分析图谱见图11，抗体偶联物7单体的纯度为96.36％。通过疏水作用高效液相色谱(HIC-HPLC)分析获得载药量(DAR)约为7-8，分析图谱见图12。Referring to the above general method, a 15-fold excess of compound (A-24) was coupled with the reduced Trastuzumab monoclonal antibody to obtain the corresponding antibody conjugate 7, and the content of the polymer was analyzed by size exclusion chromatography (SEC-HPLC). The map is shown in Figure 11, and the purity of the antibody conjugate 7 monomer was 96.36%. The drug loading (DAR) was about 7-8 by hydrophobic interaction high performance liquid chromatography (HIC-HPLC) analysis, and the analytical spectrum is shown in Figure 12.
抗体偶联物8(对照ADC3)的制备Preparation of Antibody Conjugate 8 (Control ADC3)
参照上述通用方法将12倍过量的化合物AP-1与还原后的Trastuzumab单抗偶联获得对应的抗体偶联物8，通过尺寸排阻层析(SEC-HPLC)分析聚合体的含量，分析图谱见图13，抗体偶联物7单体的纯度为97.92％。通过疏水作用高效液相色谱(HIC-HPLC)分析获得载药量(DAR)约为7-8，分析图谱见图14。Referring to the above general method, a 12-fold excess of compound AP-1 was coupled with the reduced Trastuzumab monoclonal antibody to obtain the corresponding antibody conjugate 8, and the content of the polymer was analyzed by size exclusion chromatography (SEC-HPLC), and the chromatogram was analyzed. As shown in Figure 13, the purity of the antibody conjugate 7 monomer was 97.92%. The drug loading (DAR) was about 7-8 obtained by hydrophobic interaction high performance liquid chromatography (HIC-HPLC) analysis, and the analytical spectrum is shown in Figure 14.
抗体偶联物9的制备Preparation of Antibody Conjugate 9
参照上述通用方法将12倍过量化合物(A-11)与还原后的Patritumab单抗偶联获得对应的抗体偶联物9，通过尺寸排阻层析(SEC-HPLC)分析聚合体的含量，分析图谱见图15，抗体偶联物9单体的纯度为97.92％。通过疏水作用高效液相色谱(HIC-HPLC)分析获得载药量(DAR)约为7-8，分析图谱见图16。Referring to the above general method, 12-fold excess compound (A-11) was coupled with the reduced Patritumab monoclonal antibody to obtain the corresponding antibody conjugate 9, and the content of the polymer was analyzed by size exclusion chromatography (SEC-HPLC). The map is shown in Figure 15, and the purity of the antibody conjugate 9 monomer was 97.92%. The drug loading (DAR) was about 7-8 obtained by hydrophobic interaction high performance liquid chromatography (HIC-HPLC) analysis, and the analytical spectrum is shown in Figure 16.
抗体偶联物10的制备Preparation of Antibody Conjugate 10
参照上述通用方法将12倍过量化合物(A-11)与还原后的H01L02单抗偶联获得对应的抗体偶联物10，通过尺寸排阻层析(SEC-HPLC)分析聚合体的含量，分析图谱见图17，抗体偶联物10单体的纯度为99.10％。通过疏水作用高效液相色谱(HIC-HPLC)分析获得载药量(DAR)约为7-8，分析图谱见图18。Referring to the above general method, 12 times excess compound (A-11) was coupled with the reduced H01L02 monoclonal antibody to obtain the corresponding antibody conjugate 10, and the content of the polymer was analyzed by size exclusion chromatography (SEC-HPLC). The map is shown in Figure 17, and the purity of the antibody conjugate 10 monomer was 99.10%. The drug loading (DAR) was about 7-8 by hydrophobic interaction high performance liquid chromatography (HIC-HPLC) analysis, and the analytical spectrum is shown in Figure 18.
抗体偶联物11的制备Preparation of Antibody Conjugate 11
参照上述通用方法将12倍过量化合物(A-28)与还原后的Trastuzumab单抗偶联获得对应的抗体偶联物11，通过尺寸排阻层析(SEC-HPLC)分析聚合体的含量，分析图谱见图19，抗体偶联物11单体的纯度为99.59％。通过疏水作用高效液相色谱(HIC-HPLC)分析获得载药量(DAR)约为7-8，分析图谱见图20。Referring to the above general method, a 12-fold excess of compound (A-28) was coupled with the reduced Trastuzumab monoclonal antibody to obtain the corresponding antibody conjugate 11, and the content of the polymer was analyzed by size exclusion chromatography (SEC-HPLC). The map is shown in Figure 19, and the purity of the antibody conjugate 11 monomer was 99.59%. The drug loading (DAR) was about 7-8 by hydrophobic interaction high performance liquid chromatography (HIC-HPLC) analysis, and the analytical spectrum is shown in Figure 20.
抗体偶联物12的制备Preparation of Antibody Conjugate 12
参照上述通用方法将12倍过量化合物(A-29)与还原后的Trastuzumab单抗偶联获得对应的抗体偶联物12，通过尺寸排阻层析(SEC-HPLC)分析聚合体的含量，分析图谱见图21，抗体偶联物12单体的纯度为99.67％。通过疏水作用高效液相色谱(HIC-HPLC)分析获得载药量(DAR)约为7-8，分析图谱见图22。Referring to the above general method, a 12-fold excess of compound (A-29) was coupled with the reduced Trastuzumab monoclonal antibody to obtain the corresponding antibody conjugate 12, and the content of the polymer was analyzed by size exclusion chromatography (SEC-HPLC). The map is shown in Figure 21, and the purity of the antibody conjugate 12 monomer was 99.67%. The drug loading (DAR) was about 7-8 by hydrophobic interaction high performance liquid chromatography (HIC-HPLC) analysis, and the analytical spectrum is shown in Figure 22.
抗体偶联物13的制备Preparation of Antibody Conjugate 13
将靶向Trop-2的hRS7单抗原液透析换液至50mM磷酸二氢钠-磷酸氢二钠(NaH 2PO 4-Na 2HPO 4)/50mM氯化钠(NaCl)，pH7.0缓冲液中。测量溶液中单抗浓度后用上述缓冲液稀释抗体至5mg/mL。将反应管置于冰浴中冷却降温10分钟。加入2.5倍摩尔比的三(2-羧乙基)膦盐酸盐(TCEP)，反应液于4℃搅拌过夜。在上述未纯化的反应液中继续加入适量的二甲基乙酰胺(DMA)，再分别加入6.0倍过量摩尔比的对照品药物分子或药物连接子缀合物A-11(10mM预先溶在DMA中)，保证反应体系中DMA的体积占比不超过10％，于4℃搅动2小时进行偶联。偶联结束后，向反应液中加入4.0倍摩尔比的半胱氨酸(Cysteine)小分子消耗过量的药物连接子缀合物A-11，反应于4℃搅动30分钟进行淬灭。采用脱盐柱将偶联反应混合物用pH 5.5的组氨酸-醋酸/氯化钠过滤纯化，收集滤出样品。向样品中加入十分之一体积的活性炭-组氨酸-醋酸/氯化钠的悬浊液(300mg/mL)，室温搅动2小时充分吸收游离药物小分子。然后经由0.22微米孔径的过滤装置除菌，-80℃保存。通过尺寸排阻层析(SEC-HPLC)分析聚合体的含量，分析图谱见图25，抗体偶联物13单体的纯度为98.95％。 The hRS7 single-antigen solution targeting Trop-2 was dialyzed to 50mM sodium dihydrogen phosphate-disodium hydrogen phosphate (NaH 2 PO 4 -Na 2 HPO 4 )/50mM sodium chloride (NaCl), pH7.0 buffer middle. After measuring the concentration of mAb in the solution, dilute the antibody to 5 mg/mL with the above buffer. The reaction tube was placed in an ice bath to cool down for 10 minutes. Tris(2-carboxyethyl)phosphine hydrochloride (TCEP) was added in a molar ratio of 2.5 times, and the reaction solution was stirred at 4°C overnight. Continue to add an appropriate amount of dimethylacetamide (DMA) to the above unpurified reaction solution, and then add a 6.0-fold excess molar ratio of the reference substance drug molecule or drug linker conjugate A-11 (10mM pre-dissolved in DMA) ), ensure that the volume ratio of DMA in the reaction system does not exceed 10%, and stir at 4 °C for 2 hours for coupling. After the coupling, a 4.0-fold molar ratio of Cysteine (Cysteine) small molecule was added to the reaction solution to consume the excess drug linker conjugate A-11, and the reaction was stirred at 4°C for 30 minutes for quenching. The coupling reaction mixture was purified by filtration with histidine-acetic acid/sodium chloride pH 5.5 using a desalting column, and the filtrated sample was collected. One-tenth volume of activated carbon-histidine-acetic acid/sodium chloride suspension (300 mg/mL) was added to the sample, and stirred at room temperature for 2 hours to fully absorb free small drug molecules. It was then sterilized through a 0.22 micron pore size filter and stored at -80°C. The content of the polymer was analyzed by size exclusion chromatography (SEC-HPLC), and the analysis pattern was shown in Figure 25. The purity of the antibody conjugate 13 monomer was 98.95%.
抗体-药物偶联物中的偶联药物浓度可以通过测量抗体-药物偶联物水溶液在280nm和370nm两种波长下的紫外吸光度来计算，然后进行如下计算。The conjugated drug concentration in the antibody-drug conjugate can be calculated by measuring the UV absorbance of the antibody-drug conjugate aqueous solution at two wavelengths of 280 nm and 370 nm, and then calculated as follows.
在任何给定波长处的总吸光度等于系统中所有吸收光的化学物质的吸光度之和(吸光可加和性)。因此，基于抗体与药物偶联前后抗体与药物的摩尔吸收系数不发生变化的假设，抗体-药物偶联物中抗体浓度与药物浓度由下式表示。The total absorbance at any given wavelength is equal to the sum of the absorbances of all light-absorbing chemicals in the system (absorption additivity). Therefore, based on the assumption that the molar absorption coefficient of the antibody and the drug does not change before and after conjugation of the antibody and the drug, the antibody concentration and the drug concentration in the antibody-drug conjugate are represented by the following formula.
A 280＝A D,280+A A,280＝ε D,280C D+ε A,280C A   (1) A 280 =A D,280 +A A,280 =ε D,280 C D +ε A,280 C A (1)
A 370＝A D,370+A A,370＝ε D,370C D+ε A,370C A  (2) A 370 =A D,370 +A A,370 =ε D,370 C D +ε A,370 C A (2)
其中，A 280代表抗体-药物偶联物溶液在280nm下的吸光度，A 370代表抗体-药物偶联物溶液在370nm下的吸光度。A D,280代表药物连接子缀合物在280nm下的吸光度，A D,370代表药物连接子缀合物在370nm下的吸光度。A A,280代表抗体在280nm下的吸光度，A A,370代表抗体在370nm下的吸光度。ε D,280代表药物连接子缀合物在280nm下的摩尔吸收系数，ε D,370代表药物连接子缀合物在370nm下的摩尔吸收系数，ε A,280代表抗体在280nm下的摩尔吸收系数，ε A,370代表抗体在370nm下的摩尔吸收系数。C D代表抗体-药物偶联物溶液中药物连接子缀合物的浓度，C A代表抗体-药物偶联物溶液中抗体的浓度。 Among them, A 280 represents the absorbance of the antibody-drug conjugate solution at 280 nm, and A 370 represents the absorbance of the antibody-drug conjugate solution at 370 nm. AD,280 represents the absorbance of the drug-linker conjugate at 280 nm, AD,370 represents the absorbance of the drug-linker conjugate at 370 nm. A A,280 represents the absorbance of the antibody at 280 nm, A A,370 represents the absorbance of the antibody at 370 nm. ε D,280 represents the molar absorption coefficient of the drug-linker conjugate at 280 nm, ε D,370 represents the molar absorption coefficient of the drug-linker conjugate at 370 nm, ε A,280 represents the molar absorption of the antibody at 280 nm The coefficient, ε A, 370 represents the molar absorption coefficient of the antibody at 370 nm. CD represents the concentration of the drug linker conjugate in the antibody-drug conjugate solution, and CA represents the concentration of the antibody in the antibody-drug conjugate solution.
ε A,280可以通过已知的计算方法从抗体的氨基酸序列中估计(Protein Science,1995,vol.4,2411-2423)，ε A,370一般为零。通过药物连接子缀合物在一定摩尔浓度下溶液的吸光度，根据Lambert-Beer定律，得到ε D,280和ε D,370。通过酶标仪或紫外分光光度计测出A 280和A 370的数值，并将上述4个摩尔吸收系数的值带入联立方程(1)和(2)中，可计算出C A和C D的值。则每个抗体分子中偶联药物分子的平均数目DAR＝C D/C A。化合物A-11的紫外吸光系数为ε D,280＝6480，ε D,370＝16483，并用于本例中药物抗体偶联比的计算，抗体偶联物13的DAR值约为4.0。 ε A,280 can be estimated from the amino acid sequence of the antibody by known computational methods (Protein Science, 1995, vol. 4, 2411-2423), and ε A,370 is generally zero. ε D,280 and ε D,370 are obtained according to the Lambert-Beer law by the absorbance of the solution of the drug-linker conjugate at a certain molar concentration. The values of A 280 and A 370 are measured by a microplate reader or UV spectrophotometer, and the values of the above 4 molar absorption coefficients are put into the simultaneous equations (1) and (2), and C A and C can be calculated value of D. Then the average number of conjugated drug molecules in each antibody molecule DAR=C D /C A . The UV absorption coefficients of compound A-11 are ε D,280 = 6480, ε D,370 = 16483, and used for the calculation of the drug-antibody coupling ratio in this example, the DAR value of antibody conjugate 13 is about 4.0.
抗体偶联物14的制备Preparation of Antibody Conjugate 14
将靶向Trop-2的hRS7单抗原液透析换液至50mM磷酸二氢钠-磷酸氢二钠(NaH 2PO 4-Na 2HPO 4)/50mM氯化钠(NaCl)，pH7.0缓冲液中。测量溶液中单抗浓度后用上述缓冲液稀释抗体至5mg/mL。将反应管置于冰浴中冷却降温10分钟。加入2.5倍摩尔比的三(2-羧乙基)膦盐酸盐(TCEP)，反应液于4℃搅拌过夜。在上述未纯化的反应液中继续加入适量的二甲基乙酰胺(DMA)，再分别加入6.0倍过量摩尔比的对照品药物分子或药物连接子缀合物A-14(10mM预先溶在DMA中)，保证反应体系中DMA的体积占比不超过10％，于4℃搅动2小时进行偶联。偶联结束后，向反应液中加入4.0倍摩尔比的半胱氨酸(Cysteine)小分子消耗过量的药物连接子缀合物A-14，反应于4℃搅动30分钟进行淬灭。采用脱盐柱将偶联反应混合物用pH 5.5的组氨酸-醋酸/氯化钠过滤纯化，收集滤出样品。向样品中加入十分之一体积的活性炭-组氨酸-醋酸/氯化钠的悬浊液(300mg/mL)，室温搅动2小时充分吸收游离药物小分子。然后经由0.22微米孔径的过滤装置除菌，-80℃保存。通过尺寸排阻层析(SEC-HPLC)分析聚合体的含量，分析图谱见图26，抗体偶联物14单体的纯度为98.20％。化合物A-14的紫外吸光系数为ε D,280＝5932，ε D,370＝14997，并用于本例中药物抗体偶联比的计算， 抗体偶联物14的DAR值约为4.2。 The hRS7 single-antigen solution targeting Trop-2 was dialyzed to 50mM sodium dihydrogen phosphate-disodium hydrogen phosphate (NaH 2 PO 4 -Na 2 HPO 4 )/50mM sodium chloride (NaCl), pH7.0 buffer middle. After measuring the concentration of mAb in the solution, dilute the antibody to 5 mg/mL with the above buffer. The reaction tube was placed in an ice bath to cool down for 10 minutes. Tris(2-carboxyethyl)phosphine hydrochloride (TCEP) was added in a molar ratio of 2.5 times, and the reaction solution was stirred at 4°C overnight. Continue to add an appropriate amount of dimethylacetamide (DMA) to the above unpurified reaction solution, and then add a 6.0-fold excess molar ratio of the reference substance drug molecule or drug linker conjugate A-14 (10mM pre-dissolved in DMA) ), ensure that the volume ratio of DMA in the reaction system does not exceed 10%, and stir at 4 °C for 2 hours for coupling. After the coupling, a 4.0-fold molar ratio of Cysteine was added to the reaction solution to consume the excess drug linker conjugate A-14, and the reaction was stirred at 4°C for 30 minutes for quenching. The coupling reaction mixture was purified by filtration with histidine-acetic acid/sodium chloride pH 5.5 using a desalting column, and the filtrated sample was collected. One-tenth volume of activated carbon-histidine-acetic acid/sodium chloride suspension (300 mg/mL) was added to the sample, and stirred at room temperature for 2 hours to fully absorb free small drug molecules. It was then sterilized through a 0.22 micron pore size filter and stored at -80°C. The content of the polymer was analyzed by size exclusion chromatography (SEC-HPLC), and the analysis pattern was shown in Figure 26. The purity of the antibody conjugate 14 monomer was 98.20%. The ultraviolet absorption coefficients of compound A-14 are ε D,280 =5932, ε D,370 =14997, and used for the calculation of the drug-antibody coupling ratio in this example, the DAR value of the antibody conjugate 14 is about 4.2.
抗体偶联物15的制备Preparation of Antibody Conjugate 15
将靶向Trop-2的hRS7单抗原液透析换液至50mM磷酸二氢钠-磷酸氢二钠(NaH 2PO 4-Na 2HPO 4)/50mM氯化钠(NaCl)，pH7.0缓冲液中。测量溶液中单抗浓度后用上述缓冲液稀释抗体至5mg/mL。将反应管置于冰浴中冷却降温10分钟。加入2.5倍摩尔比的三(2-羧乙基)膦盐酸盐(TCEP)，反应液于4℃搅拌过夜。在上述未纯化的反应液中继续加入适量的二甲基乙酰胺(DMA)，再分别加入6.0倍过量摩尔比的对照品药物分子或药物连接子缀合物A-24(10mM预先溶在DMA中)，保证反应体系中DMA的体积占比不超过10％，于4℃搅动2小时进行偶联。偶联结束后，向反应液中加入4.0倍摩尔比的半胱氨酸(Cysteine)小分子消耗过量的药物连接子缀合物A-24，反应于4℃搅动30分钟进行淬灭。采用脱盐柱将偶联反应混合物用pH 5.5的组氨酸-醋酸/氯化钠过滤纯化，收集滤出样品。向样品中加入十分之一体积的活性炭-组氨酸-醋酸/氯化钠的悬浊液(300mg/mL)，室温搅动2小时充分吸收游离药物小分子。然后经由0.22微米孔径的过滤装置除菌，-80℃保存。通过尺寸排阻层析(SEC-HPLC)分析聚合体的含量，分析图谱见图27，抗体偶联物15单体的纯度为96.36％。化合物A-24的紫外吸光系数为ε D,280＝4723，ε D,370＝12467，并用于本例中药物抗体偶联比的计算，抗体偶联物15的DAR值约为4.0。 The hRS7 single-antigen solution targeting Trop-2 was dialyzed to 50mM sodium dihydrogen phosphate-disodium hydrogen phosphate (NaH 2 PO 4 -Na 2 HPO 4 )/50mM sodium chloride (NaCl), pH7.0 buffer middle. After measuring the concentration of mAb in the solution, dilute the antibody to 5 mg/mL with the above buffer. The reaction tube was placed in an ice bath to cool down for 10 minutes. Tris(2-carboxyethyl)phosphine hydrochloride (TCEP) was added in a molar ratio of 2.5 times, and the reaction solution was stirred at 4°C overnight. Continue to add an appropriate amount of dimethylacetamide (DMA) to the above unpurified reaction solution, and then add a 6.0-fold excess molar ratio of the reference substance drug molecule or drug linker conjugate A-24 (10mM pre-dissolved in DMA) ), ensure that the volume ratio of DMA in the reaction system does not exceed 10%, and stir at 4 °C for 2 hours for coupling. After the coupling was completed, 4.0 times the molar ratio of Cysteine (Cysteine) small molecule was added to the reaction solution to consume the excess drug linker conjugate A-24, and the reaction was stirred at 4°C for 30 minutes for quenching. The coupling reaction mixture was purified by filtration with histidine-acetic acid/sodium chloride pH 5.5 using a desalting column, and the filtrated sample was collected. One-tenth volume of activated carbon-histidine-acetic acid/sodium chloride suspension (300 mg/mL) was added to the sample, and stirred at room temperature for 2 hours to fully absorb free small drug molecules. It was then sterilized through a 0.22 micron pore size filter and stored at -80°C. The content of the polymer was analyzed by size exclusion chromatography (SEC-HPLC), and the analysis pattern was shown in Figure 27. The purity of the antibody conjugate 15 monomer was 96.36%. The UV absorption coefficients of compound A-24 are ε D,280 = 4723, ε D,370 = 12467, and used for the calculation of the drug-antibody coupling ratio in this example, the DAR value of antibody conjugate 15 is about 4.0.
本申请的化合物以及其制备的ADC对比对照分子制备的ADC分子在更高载药量的情况下单体纯度更高，提示该类ADC分子具有更佳的稳定性。此外，疏水作用高效液相色谱(HIC-HPLC)分析结果显示该本申请的ADC分子较对照分子有更低的保留时间，与祼抗更接近，提示该类ADC分子具有更佳的亲水性，在体内可能具有更长的半衰期与更强的体内药效。Compared with the ADC molecules prepared by the compounds of the present application and the ADC prepared by the control molecules, the monomer purity is higher under the condition of higher drug loading, suggesting that such ADC molecules have better stability. In addition, the results of hydrophobic interaction high performance liquid chromatography (HIC-HPLC) analysis showed that the ADC molecule of the present application had a lower retention time than the reference molecule, and was closer to the naked antibody, suggesting that this type of ADC molecule has better hydrophilicity , may have a longer half-life and stronger in vivo efficacy.
实施例3抗体偶联物稳定性实验测试Example 3 Antibody Conjugate Stability Experimental Test
基于喜树碱类拓扑异构酶抑制剂的ADC药物如Sacituzumab govitecan(Trodelvy)与Trastuzumab Deruxtecan(Enhertu)通常情况下具有较高的药物抗体荷载比(DAR)，由于喜树碱类分子具有芳香稠环类结构，具有较强的疏水性性，具得该类ADC分子的稳定性必然受到一定影响。本发明所制备的ADC在特定位置引入了聚肌氨酸肽链，具有更好的“屏障效应”，可能具有较好的稳定性，我们设计了抗体稳定性加速试验来验证这一假设。ADC drugs based on camptothecin-like topoisomerase inhibitors such as Sacituzumab govitecan (Trodelvy) and Trastuzumab Deruxtecan (Enhertu) usually have a higher drug-to-antibody loading ratio (DAR), due to the aromatic concentration of camptothecin-like molecules. The ring-like structure has strong hydrophobicity, and the stability of such ADC molecules must be affected to a certain extent. The ADC prepared by the present invention introduces a polysarcosine peptide chain at a specific position, which has better "barrier effect" and may have better stability. We designed an antibody stability accelerated test to verify this hypothesis.
比较的分子包括：Molecules compared include:
1.曲妥珠单抗1. Trastuzumab
2.抗体偶联物22. Antibody Conjugate 2
3.抗体偶联物53. Antibody Conjugate 5
4.抗体偶联物64. Antibody Conjugate 6
5.抗体偶联物75. Antibody Conjugate 7
5.抗体偶联物85. Antibody Conjugates 8
我们将待测分子样品用制剂缓冲液(20mM Histidine-acetic acid,150mM NaCl,pH 5.5)稀释至浓度为5毫克/毫升，取50μL直接进行测试或放置于55℃水浴锅中分别孵育1小时、2小时、5小时、24小时、48小时与72小时之后再进行测试(稳定性加速实验)。分别通过UV法检测样品浓度变化，以及在安捷伦1260 Infinity II生物惰性液相色谱系统上采用TSK-GEL SWXL3000分子排阻色谱柱对样品中的正常结构ADC分子(主峰)、聚合分子的含量进行定量分析。We diluted the molecular samples to be tested with preparation buffer (20mM Histidine-acetic acid, 150mM NaCl, pH 5.5) to a concentration of 5 mg/ml, and took 50 μL for testing directly or placed them in a water bath at 55°C for 1 hour, Tests were performed again after 2 hours, 5 hours, 24 hours, 48 hours and 72 hours (stability acceleration test). The change of sample concentration was detected by UV method, and the content of normal structure ADC molecules (main peak) and polymeric molecules in the sample was quantified by TSK-GEL SWXL3000 size exclusion chromatography column on Agilent 1260 Infinity II bio-inert liquid chromatography system. analyze.
HPLC的流动相为200mM磷酸盐缓冲液(pH 7.0)+150mM KCl+15％IPA，柱温25℃，进样体积为7μL，流速为0.75毫升/分钟，UV检测波长280nm和370nm。The mobile phase of HPLC was 200 mM phosphate buffer (pH 7.0) + 150 mM KCl + 15% IPA, the column temperature was 25 °C, the injection volume was 7 μL, the flow rate was 0.75 ml/min, and the UV detection wavelengths were 280 nm and 370 nm.
样品浓度变化统计结果如图23所示，我们发现曲妥珠单抗与其对应的抗体偶联药物5、抗体偶联药物6均具有较好的热稳定性，0-72小时过程中浓度变化差异较小，而抗体偶联药物2、抗体偶联药物7与抗体偶联药物8的样品在24小时后出现ADC浓度快速下降并发生沉淀现象，从而导致样品浓度快速下降。The statistical results of sample concentration changes are shown in Figure 23. We found that trastuzumab and its corresponding antibody-drug conjugates 5 and antibody-drug conjugates 6 have good thermal stability, and the concentration changes during 0-72 hours are different. However, the samples of antibody conjugated drug 2, antibody conjugated drug 7 and antibody conjugated drug 8 showed a rapid decrease in ADC concentration and precipitation after 24 hours, resulting in a rapid decrease in sample concentration.
进一步通过分子排阻色谱对上述加速稳定性实验样品中的聚合分子增加比例变化结果进行分析，统计结果如图24所示，与样品浓度变化趋势一致，样品稳定性从高到低排序为曲妥珠单抗≈抗体偶联物5>抗体偶联物6>抗体偶联物7>抗体偶联物8≈抗体偶联物2。The results of the increase in the proportion of polymer molecules in the accelerated stability test samples were further analyzed by size exclusion chromatography. The statistical results are shown in Figure 24, which is consistent with the trend of sample concentration changes. The stability of the samples is ranked from high to low. Bezumab ≈ Antibody Conjugate 5 > Antibody Conjugate 6 > Antibody Conjugate 7 > Antibody Conjugate 8 ≈ Antibody Conjugate 2.
上述结果验证了本发明特定位置的亲水性聚肌氨酸肽链具有“屏障效应”为抗体药物偶联物分子带来了“意想不到”的稳定性提高的效果。The above results verify that the hydrophilic polysarcosine peptide chain at a specific position of the present invention has a "barrier effect", which brings an "unexpected" effect of improving the stability of the antibody drug conjugate molecule.
实施例4体外细胞毒活性测试Example 4 In vitro cytotoxic activity test
选择人肺鳞癌细胞NCI-H2170与LK-2，人乳腺癌细胞MDA-MB-231作为本次实验体外活性检测用细胞株，通过流式细胞技术(FACS)测定上述人肿瘤细胞表面TROP2抗原表达量情况(通过rMFI表示)，同时观察抗体偶联物13、DS-1062a(对照药物，参照US11173213B2中描述方法进行制备)、hIgG-deruxtecan偶联物、hIgG-A-11偶联物对细胞杀伤的量效情况。抗体偶联药物加样后终浓度设定500nM为起始浓度，500～0.1nM设计系列 9个浓度(5倍比稀释)，观察120小时的杀伤(或抑制)变化，化学发光染色(Luminescent Cell Viability Assay)，读取荧光数据后计算IC 50。 Human lung squamous cell carcinoma cells NCI-H2170 and LK-2, and human breast cancer cell MDA-MB-231 were selected as the cell lines for in vitro activity detection in this experiment, and the surface TROP2 antigen of the above human tumor cells was determined by flow cytometry (FACS). The expression level (expressed by rMFI), and the effect of antibody conjugate 13, DS-1062a (control drug, prepared according to the method described in US11173213B2), hIgG-deruxtecan conjugate, and hIgG-A-11 conjugate on cells was observed at the same time. The dose-response situation of killing. The final concentration of the antibody conjugated drug after adding the sample was set at 500nM as the initial concentration, and 500-0.1nM was designed in a series of 9 concentrations (5-fold ratio dilution), and the killing (or inhibition) changes were observed for 120 hours, and chemiluminescence staining (Luminescent Cell Viability Assay), IC 50 was calculated after reading the fluorescence data.
如表1所示，抗体偶联物13、DS-1062a(对照药物)均能明显抑制肿瘤细胞增殖，显著强于阴性对阳药物IgG-deruxtecan偶联物、IgG A-1偶联物，本发明的抗体偶联物13明显优于临床药物DS-1062a。As shown in Table 1, both antibody conjugate 13 and DS-1062a (control drug) can significantly inhibit the proliferation of tumor cells, which are significantly stronger than the negative and positive drugs IgG-deruxtecan conjugate and IgG A-1 conjugate. The inventive antibody conjugate 13 is significantly better than the clinical drug DS-1062a.
表1Table 1
Figure PCTCN2022089726-appb-000107
Figure PCTCN2022089726-appb-000107
综合以上活性测试结果看，本申请的化合物制备得的ADC均表现良好的体外抗肿瘤活性，特别对于相关抗原低表达的肿瘤细胞本申请的抗体偶联物优势更为明显。Based on the above activity test results, the ADCs prepared from the compounds of the present application all show good in vitro anti-tumor activity, especially for tumor cells with low expression of related antigens. The antibody conjugates of the present application have more obvious advantages.
实施例5体内抗肿瘤功效测定Example 5 Determination of anti-tumor efficacy in vivo
可以在体内测量本发明的组合的功效，即在啮齿类动物中植入癌细胞的同种异体移植物或异种移植物，并用所述组合处理肿瘤。将受试小鼠用药物或对照处理，并监测数周或更长时间以测量到达肿瘤倍增的时间、对数细胞杀伤和肿瘤抑制。The efficacy of the combinations of the invention can be measured in vivo, ie in rodents implanted with cancer cell allografts or xenografts, and treated with the combination. Test mice are treated with drug or control and monitored for several weeks or longer to measure time to tumor doubling, log cell killing, and tumor inhibition.
体内抗肿瘤实验In vivo antitumor experiments
将作为人肺鳞癌细胞NCI-H2170(ATCC)悬浮于生理盐水，将4×10 7个细胞向雌性裸鼠的右体侧部皮下移植，在第6天随机实施分组。将分组日作为第0天，在第0天，分别以5mg/kg的用量向尾静脉内给予抗体偶联物13，DS-1062a(阳性对照药物)、hIgG-A-11偶联物作为阴性对照组。 Human lung squamous cell carcinoma cell NCI-H2170 (ATCC) was suspended in physiological saline, 4×10 7 cells were subcutaneously transplanted into the right flank of female nude mice, and grouping was performed at random on the sixth day. Taking the grouping day as the 0th day, on the 0th day, the antibody conjugate 13, DS-1062a (positive control drug) and hIgG-A-11 conjugate were respectively administered to the tail vein at a dose of 5 mg/kg as the negative control group.
结果示于图28，对于人肺鳞癌细胞NCI-H2170，给予5mg/kg的DS-1062a、抗体偶联物13均显示出抗肿瘤活性，5mg/kg单次给药剂量下，抗体偶联物13的抑瘤活性显著强于对照药物DS-1062a。此外从小鼠体重变化趋势来看(图29)，提示抗体偶联物13安全性良好。The results are shown in Figure 28. For human lung squamous cell carcinoma NCI-H2170, DS-1062a and antibody conjugate 13 at 5 mg/kg both showed anti-tumor activity. At a single dose of 5 mg/kg, the antibody conjugated The antitumor activity of compound 13 was significantly stronger than that of the control drug DS-1062a. In addition, from the trend of changes in the body weight of mice (Fig. 29), it was suggested that the antibody conjugate 13 was safe.
从活性测试结果看，本申请的化合物制备得的ADC均表现出一定的体内抗肿瘤活性，与对照样品相比可以显示出显著更强的抗肿瘤活性。荷瘤小鼠对以上药物均能很好耐受，没 有体重减轻等症状发生。From the activity test results, the ADCs prepared from the compounds of the present application all showed a certain anti-tumor activity in vivo, and could show significantly stronger anti-tumor activity compared with the control sample. The tumor-bearing mice tolerated the above drugs well, and no symptoms such as weight loss occurred.
前述详细说明是以解释和举例的方式提供的，并非要限制所附权利要求的范围。目前本申请所列举的实施方式的多种变化对本领域普通技术人员来说是显而易见的，且保留在所附的权利要求和其等同方案的范围内。The foregoing detailed description has been presented by way of explanation and example, and is not intended to limit the scope of the appended claims. Various modifications to the embodiments presently enumerated in this application will be apparent to those of ordinary skill in the art and remain within the scope of the appended claims and their equivalents.

Claims (43)

  1. 一种化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，其中所述化合物包含式(C-Trop-2)所示的结构：A compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, pro- A drug or solvate, wherein the compound comprises a structure represented by the formula (C-Trop-2):
    Figure PCTCN2022089726-appb-100001
    Figure PCTCN2022089726-appb-100001
    其中，Q 1包含连接体， where Q1 contains the linker,
    L 1包含-L 1a-C(＝O)-， L 1 contains -L 1a -C(=O)-,
    其中，L 1a选自以下组：任选取代的亚烷基、任选取代的聚乙二醇基、任选取代的亚烯基、任选取代的亚炔基、任选取代的亚脂环基、任选取代的亚脂杂环基、任选取代的亚芳基和任选取代的亚杂芳基； wherein, L 1a is selected from the group consisting of optionally substituted alkylene, optionally substituted polyethylene glycol, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted alicyclic groups, optionally substituted alicyclic heterocyclyl groups, optionally substituted arylene groups, and optionally substituted heteroarylene groups;
    L 2包含任选取代的多肽残基， L2 comprises optionally substituted polypeptide residues,
    L 3包含任选取代的间隔基团， L 3 comprises an optionally substituted spacer group,
    其中，L 2和/或L 3包含任选取代的聚肌氨酸残基， wherein L 2 and/or L 3 comprise optionally substituted polysarcosine residues,
    T包含药物单元，T contains the drug unit,
    Ab为能够结合Trop-2的配体，m为1-8的数。Ab is a ligand capable of binding Trop-2, and m is a number from 1 to 8.
  2. 根据权利要求1所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of claim 1, or a tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable form thereof an accepted salt, prodrug or solvate,
    其中，Q 1包含与巯基偶联后的连接体。 Wherein, Q 1 includes a linker coupled with a thiol group.
  3. 根据权利要求1-2中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-2, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，Q 1选自以下组：任选取代的
    Figure PCTCN2022089726-appb-100002
    任选取代的
    Figure PCTCN2022089726-appb-100003
    任选取代的
    Figure PCTCN2022089726-appb-100004
    和任选取代的
    Figure PCTCN2022089726-appb-100005
    wherein Q 1 is selected from the group consisting of: optionally substituted
    Figure PCTCN2022089726-appb-100002
    optionally substituted
    Figure PCTCN2022089726-appb-100003
    optionally substituted
    Figure PCTCN2022089726-appb-100004
    and optionally substituted
    Figure PCTCN2022089726-appb-100005
  4. 根据权利要求1-3中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-3, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，L 1a选自以下组：任选取代的C 1-C 7的亚烷基、任选取代的二甘醇基至八甘醇基、任选取代的C 3-C 6亚脂环基、任选取代的亚芳基和任选取代的亚杂芳基。 wherein, L 1a is selected from the group consisting of optionally substituted C 1 -C 7 alkylene groups, optionally substituted diethylene glycol to octaethylene glycol groups, optionally substituted C 3 -C 6 alicyclic groups , optionally substituted arylene and optionally substituted heteroarylene.
  5. 根据权利要求1-4中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-4, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，L 1a选自以下组：任选取代的亚甲基、任选取代的亚乙基、任选取代的亚丙基、任选取代的亚丁基、任选取代的亚戊基、任选取代的二甘醇基、任选取代的四甘醇基、任选取代的六甘醇基、任选取代的八甘醇基和任选取代的亚环己基。 wherein L 1a is selected from the group consisting of optionally substituted methylene, optionally substituted ethylene, optionally substituted propylene, optionally substituted butylene, optionally substituted pentylene, optionally substituted Substituted diethylene glycol, optionally substituted tetraethylene glycol, optionally substituted hexaethylene glycol, optionally substituted octaethylene glycol, and optionally substituted cyclohexylene.
  6. 根据权利要求1-5中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-5, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，L 2包含任选取代的选自以下组的氨基酸构成的多肽残基：苯丙氨酸、异亮氨酸、亮氨酸、色氨酸、缬氨酸、甲硫氨酸、酪氨酸、丙氨酸、苏氨酸、组氨酸、丝氨酸、谷氨酰胺、精氨酸、赖氨酸、天冬酰胺、谷氨酸、脯氨酸、瓜氨酸、天冬氨酸和甘氨酸。 wherein L 2 comprises optionally substituted polypeptide residues consisting of amino acids selected from the group consisting of phenylalanine, isoleucine, leucine, tryptophan, valine, methionine, tyrosine acid, alanine, threonine, histidine, serine, glutamine, arginine, lysine, asparagine, glutamic acid, proline, citrulline, aspartic acid and glycine .
  7. 根据权利要求1-6中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-6, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，L 2包含任选取代的选自以下组的氨基酸构成的多肽残基：甘氨酸、苯丙氨酸、缬氨酸、丙氨酸、精氨酸、瓜氨酸、天冬氨酸、天冬酰胺和赖氨酸。 wherein L 2 comprises optionally substituted polypeptide residues consisting of amino acids selected from the group consisting of: glycine, phenylalanine, valine, alanine, arginine, citrulline, aspartic acid, amino acid Paraparagine and Lysine.
  8. 根据权利要求1-7中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-7, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，L 2包含任选取代的选自以下组的多肽残基：苯丙氨酸-赖氨酸(Phe-Lys)、缬氨酸-丙氨酸(Val-Ala)、缬氨酸-瓜氨酸(Val-Cit)、谷氨酸-缬氨酸-丙氨酸(Glu-Val-Ala)、谷氨酸-缬氨酸-瓜氨酸(Glu-Val-Cit)、缬氨酸-赖氨酸(Val-Lys)、丙氨酸-丙氨酸-丙氨酸(Ala-Ala-Ala)、丙氨酸-丙氨酸-天冬酰胺(Ala-Ala-Asn)和甘氨酸-甘氨酸-苯丙氨酸-甘氨酸(Gly-Gly-Phe-Gly)。 wherein L 2 comprises an optionally substituted polypeptide residue selected from the group consisting of phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala), valine-citrulline amino acid (Val-Cit), glutamic acid-valine-alanine (Glu-Val-Ala), glutamic acid-valine-citrulline (Glu-Val-Cit), valine- Lysine (Val-Lys), Alanine-Alanine-Alanine (Ala-Ala-Ala), Alanine-Alanine-Asparagine (Ala-Ala-Asn), and Glycine-Glycine - Phenylalanine-Glycine (Gly-Gly-Phe-Gly).
  9. 根据权利要求1-8中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-8, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，当L 2包含赖氨酸残基时，所述赖氨酸残基被包含聚肌氨酸残基的结构R 1取代。 Wherein, when L2 contains a lysine residue, the lysine residue is replaced by a structure R1 containing a polysarcosine residue.
  10. 根据权利要求9所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构 体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of claim 9, or a tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable form thereof an accepted salt, prodrug or solvate,
    其中，所述R 1为任选取代的
    Figure PCTCN2022089726-appb-100006
    其中n1为4至18的数，R选自以下组：C 1-C 6烷基、C 1-C 6环烷基、和C 1-C 6烷氧基。     wherein, the R 1 is optionally substituted
    Figure PCTCN2022089726-appb-100006
    wherein n1 is a number from 4 to 18, and R is selected from the group consisting of C 1 -C 6 alkyl, C 1 -C 6 cycloalkyl, and C 1 -C 6 alkoxy.
  11. 根据权利要求10所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of claim 10, or a tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable form thereof an accepted salt, prodrug or solvate,
    其中，n1为4至18。where n1 is 4 to 18.
  12. 根据权利要求1-11中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-11, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，L 3选自以下组：任选取代的
    Figure PCTCN2022089726-appb-100007
    和任选取代的
    Figure PCTCN2022089726-appb-100008
    wherein L is selected from the group consisting of optionally substituted
    Figure PCTCN2022089726-appb-100007
    and optionally substituted
    Figure PCTCN2022089726-appb-100008
  13. 根据权利要求1-12中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-12, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，L 3的苯环与任选取代的聚肌氨酸残基通过结构单元-X-连接，结构单元-X-选自以下组：任选取代的
    Figure PCTCN2022089726-appb-100009
    其中X 1选自以下组：羰基、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；其中X 2选自以下组：氢、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；其中X 3为共价键或选自以下组：氢、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；所述的C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基各自独立地任选地被选自氘代、卤素、氰基、硝基、氨基、烷基、羧基、烷氧基或环烷基的一个或者多个取代基取代。     Wherein, the benzene ring of L 3 is connected with the optionally substituted polysarcosine residue through the structural unit -X-, and the structural unit -X- is selected from the following group: optionally substituted
    Figure PCTCN2022089726-appb-100009
    wherein X 1 is selected from the group consisting of carbonyl, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight chain heteroalkyl containing 1-8 atoms, and Linear-cyclic heteroalkyl groups containing 1-8 atoms containing 1-3 atoms selected from N, O or S ; wherein X is selected from the group consisting of hydrogen, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkane containing 1-8 atoms group, the heteroalkyl group contains 1-3 atoms selected from N, O or S; wherein X 3 is a covalent bond or is selected from the following group: hydrogen, C 1 -C 8 alkyl, C 1 -C 8 Alkoxy, C1 - C6cycloalkyl, straight-chain heteroalkyl containing 1-8 atoms, and straight-cyclic-cyclic heteroalkyl containing 1-8 atoms, the heteroalkyl containing selected 1-3 atoms from N, O or S; said C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight containing 1-8 atoms Chain heteroalkyl, and straight-chain-cyclic heteroalkyl containing 1-8 atoms are each independently optionally selected from deuterated, halogen, cyano, nitro, amino, alkyl, carboxy, alkoxy substituted with one or more substituents of the radical or cycloalkyl.
  14. 根据权利要求1-13中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合 物，The compound of any one of claims 1-13, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，L 3的苯环与任选取代的聚肌氨酸残基通过结构单元-X-连接，结构单元-X-选自以下组：任选取代的
    Figure PCTCN2022089726-appb-100010
    和任选取代的
    Figure PCTCN2022089726-appb-100011
    其中X 1选自以下组：C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子，所述的C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基各自独立地任选地选自氘代、卤素、氰基、硝基、氨基、烷基、羧基、烷氧基或环烷基的一个或者多个取代基取代。     Wherein, the benzene ring of L 3 is connected with the optionally substituted polysarcosine residue through the structural unit -X-, and the structural unit -X- is selected from the following group: optionally substituted
    Figure PCTCN2022089726-appb-100010
    and optionally substituted
    Figure PCTCN2022089726-appb-100011
    wherein X 1 is selected from the group consisting of C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight chain heteroalkyl containing 1-8 atoms, and containing 1 -8-atom straight-chain-cyclic heteroalkyl, said heteroalkyl containing 1-3 atoms selected from N, O or S, said C 1 -C 8 alkyl, C 1 -C 8 Alkoxy, C1 - C6cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkyl containing 1-8 atoms are each independently optionally selected from Substituted with one or more substituents of deuterated, halogen, cyano, nitro, amino, alkyl, carboxyl, alkoxy or cycloalkyl.
  15. 根据权利要求13-14中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 13-14, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，结构单元-X-为任选取代的
    Figure PCTCN2022089726-appb-100012
    所述任选取代的聚肌氨酸残基包含
    Figure PCTCN2022089726-appb-100013
    其中n2为4至18的数，R选自以下组：C 1-C 6烷基、C 1-C 6环烷基、和C 1-C 6烷氧基。     wherein the structural unit -X- is optionally substituted
    Figure PCTCN2022089726-appb-100012
    The optionally substituted polysarcosine residue comprises
    Figure PCTCN2022089726-appb-100013
    wherein n2 is a number from 4 to 18, and R is selected from the group consisting of C 1 -C 6 alkyl, C 1 -C 6 cycloalkyl, and C 1 -C 6 alkoxy.
  16. 根据权利要求13-15中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 13-15, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，结构单元-X-选自以下组：任选取代的
    Figure PCTCN2022089726-appb-100014
    其中X 1选自以下组：羰基、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；其中X 2选自以下组：氢、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；其中X 3为共价键或选自以下组：氢、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；所述的C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基各自独立地任选地被选自氘代、卤素、氰基、硝基、氨 基、烷基、羧基、烷氧基或环烷基的一个或者多个取代基取代，所述任选取代的聚肌氨酸残基包含
    Figure PCTCN2022089726-appb-100015
    其中n2为4至18的数，R选自以下组：C 1-C 6烷基、C 1-C 6环烷基、和C 1-C 6烷氧基。     wherein the structural unit -X- is selected from the group consisting of: optionally substituted
    Figure PCTCN2022089726-appb-100014
    wherein X 1 is selected from the group consisting of carbonyl, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight chain heteroalkyl containing 1-8 atoms, and Linear-cyclic heteroalkyl groups containing 1-8 atoms containing 1-3 atoms selected from N, O or S ; wherein X is selected from the group consisting of hydrogen, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkane containing 1-8 atoms group, the heteroalkyl group contains 1-3 atoms selected from N, O or S; wherein X 3 is a covalent bond or is selected from the following group: hydrogen, C 1 -C 8 alkyl, C 1 -C 8 Alkoxy, C1 - C6cycloalkyl, straight-chain heteroalkyl containing 1-8 atoms, and straight-cyclic-cyclic heteroalkyl containing 1-8 atoms, the heteroalkyl containing selected 1-3 atoms from N, O or S; said C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight containing 1-8 atoms Chain heteroalkyl, and straight-chain-cyclic heteroalkyl containing 1-8 atoms are each independently optionally selected from deuterated, halogen, cyano, nitro, amino, alkyl, carboxy, alkoxy substituted with one or more substituents of a radical or cycloalkyl, the optionally substituted polysarcosine residue comprising
    Figure PCTCN2022089726-appb-100015
    wherein n2 is a number from 4 to 18, and R is selected from the group consisting of C 1 -C 6 alkyl, C 1 -C 6 cycloalkyl, and C 1 -C 6 alkoxy.
  17. 根据权利要求13-14中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 13-14, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，结构单元-X-为任选取代的
    Figure PCTCN2022089726-appb-100016
    所述任选取代的聚肌氨酸残基包含
    Figure PCTCN2022089726-appb-100017
    其中n2为4至18的数，R选自以下组：C 1-C 6烷基、C 1-C 6环烷基、和C 1-C 6烷氧基。     wherein the structural unit -X- is optionally substituted
    Figure PCTCN2022089726-appb-100016
    The optionally substituted polysarcosine residue comprises
    Figure PCTCN2022089726-appb-100017
    wherein n2 is a number from 4 to 18, and R is selected from the group consisting of C 1 -C 6 alkyl, C 1 -C 6 cycloalkyl, and C 1 -C 6 alkoxy.
  18. 根据权利要求13-14和17中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 13-14 and 17, or a tautomer, meso, racemate, enantiomer, diastereomer, or tautomer thereof in the form of a mixture, or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，结构单元-X-选自以下组：任选取代的
    Figure PCTCN2022089726-appb-100018
    其中X 1选自以下组：羰基、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；其中X 2选自以下组：氢、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；其中X 3为共价键或选自以下组：氢、C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基，所述杂烷基包含选自N、O或者S的1-3个原子；所述的C 1-C 8烷基、C 1-C 8烷氧基、C 1-C 6环烷基、包含1-8个原子的直链杂烷基、和包含1-8个原子的直链-环状杂烷基各自独立地任选地被选自氘代、卤素、氰基、硝基、氨基、烷基、羧基、烷氧基或环烷基的一个或者多个取代基取代，所述任选取代的聚肌氨酸残基包含
    Figure PCTCN2022089726-appb-100019
    其中n2为4至18的数，R选自以下组：C 1-C 6烷基、C 1-C 6环烷基、 和C 1-C 6烷氧基。     wherein the structural unit -X- is selected from the group consisting of: optionally substituted
    Figure PCTCN2022089726-appb-100018
    wherein X 1 is selected from the group consisting of carbonyl, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight chain heteroalkyl containing 1-8 atoms, and Linear-cyclic heteroalkyl groups containing 1-8 atoms containing 1-3 atoms selected from N, O or S ; wherein X is selected from the group consisting of hydrogen, C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, linear heteroalkyl containing 1-8 atoms, and linear-cyclic heteroalkane containing 1-8 atoms group, the heteroalkyl group contains 1-3 atoms selected from N, O or S; wherein X 3 is a covalent bond or is selected from the following group: hydrogen, C 1 -C 8 alkyl, C 1 -C 8 Alkoxy, C1 - C6cycloalkyl, straight-chain heteroalkyl containing 1-8 atoms, and straight-cyclic-cyclic heteroalkyl containing 1-8 atoms, the heteroalkyl containing selected 1-3 atoms from N, O or S; said C 1 -C 8 alkyl, C 1 -C 8 alkoxy, C 1 -C 6 cycloalkyl, straight containing 1-8 atoms Chain heteroalkyl, and straight-chain-cyclic heteroalkyl containing 1-8 atoms are each independently optionally selected from deuterated, halogen, cyano, nitro, amino, alkyl, carboxy, alkoxy substituted with one or more substituents of a radical or cycloalkyl, the optionally substituted polysarcosine residue comprising
    Figure PCTCN2022089726-appb-100019
    wherein n2 is a number from 4 to 18, and R is selected from the group consisting of C 1 -C 6 alkyl, C 1 -C 6 cycloalkyl, and C 1 -C 6 alkoxy.
  19. 根据权利要求15-18中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 15-18, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，n2为4至18。where n2 is 4 to 18.
  20. 根据权利要求1-19中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-19, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，T包含具有抗肿瘤活性的化合物。Wherein, T comprises a compound with antitumor activity.
  21. 根据权利要求1-20中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-20, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，T包含拓扑异构酶抑制剂。where T comprises a topoisomerase inhibitor.
  22. 根据权利要求1-21中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-21, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，T包含喜树碱类及非喜树碱类拓扑异构酶I抑制剂。Wherein, T includes camptothecin and non-camptothecin topoisomerase I inhibitors.
  23. 根据权利要求1-22中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-22, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中，T选自以下组的结构：
    Figure PCTCN2022089726-appb-100020
    where T is selected from the structure of the following group:
    Figure PCTCN2022089726-appb-100020
  24. 根据权利要求1-23中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-23, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中所述Ab包含抗Trop-2抗体或其抗原结合片段。wherein the Ab comprises an anti-Trop-2 antibody or an antigen-binding fragment thereof.
  25. 根据权利要求24所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of claim 24, or a tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable form thereof an accepted salt, prodrug or solvate,
    其中所述抗体选自以下组：鼠源抗体、嵌合抗体、人源化抗体和全人源抗体。wherein the antibody is selected from the group consisting of murine antibodies, chimeric antibodies, humanized antibodies and fully human antibodies.
  26. 根据权利要求24-25中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 24-25, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中所述抗体包含单克隆抗体。wherein the antibody comprises a monoclonal antibody.
  27. 根据权利要求24-26中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 24-26, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中所述抗体包含双特异性抗体。wherein the antibody comprises a bispecific antibody.
  28. 根据权利要求24-27中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 24-27, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中所述抗原结合片段选自以下组：Fab，Fab′，Fv片段，F(ab') 2，F(ab) 2，scFv，di-scFv，VHH和dAb。 wherein the antigen-binding fragment is selected from the group consisting of Fab, Fab', Fv fragments, F(ab') 2 , F(ab) 2 , scFv, di-scFv, VHH and dAb.
  29. 根据权利要求1-28中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-28, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中所述Ab的重链HCDR1、HCDR2和HCDR3和轻链LCDR1、LCDR2和LCDR3分别包含抗Trop-2抗体的重链HCDR1、HCDR2和HCDR3和轻链LCDR1、LCDR2和LCDR3。Wherein the heavy chain HCDR1, HCDR2 and HCDR3 of the Ab and the light chain LCDR1, LCDR2 and LCDR3 respectively comprise the heavy chain HCDR1, HCDR2 and HCDR3 and the light chain LCDR1, LCDR2 and LCDR3 of the anti-Trop-2 antibody.
  30. 根据权利要求1-29中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-29, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中所述Ab的重链HCDR1、HCDR2和HCDR3和轻链LCDR1、LCDR2和LCDR3分别包含hRS7的重链HCDR1、HCDR2和HCDR3和轻链LCDR1、LCDR2和LCDR3。Wherein the heavy chain HCDR1, HCDR2 and HCDR3 and the light chain LCDR1, LCDR2 and LCDR3 of the Ab respectively comprise the heavy chain HCDR1, HCDR2 and HCDR3 and the light chain LCDR1, LCDR2 and LCDR3 of hRS7.
  31. 根据权利要求1-30中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-30, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中所述Ab的重链可变区VH和轻链可变区VL分别包含抗Trop-2抗体的重链可变区VH和轻链可变区VL。The heavy chain variable region VH and the light chain variable region VL of the Ab respectively comprise the heavy chain variable region VH and the light chain variable region VL of the anti-Trop-2 antibody.
  32. 根据权利要求1-31中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-31, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中所述Ab的重链可变区VH和轻链可变区VL分别包含hRS7的重链可变区VH和轻链可变区VL。The heavy chain variable region VH and the light chain variable region VL of the Ab respectively comprise the heavy chain variable region VH and the light chain variable region VL of hRS7.
  33. 根据权利要求1-32中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-32, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中所述Ab的重链和轻链分别包含抗Trop-2抗体的重链和轻链。wherein the heavy chain and light chain of the Ab comprise the heavy chain and light chain of an anti-Trop-2 antibody, respectively.
  34. 根据权利要求1-33中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-33, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中所述Ab的重链和轻链分别包含hRS7的重链和轻链。wherein the heavy chain and light chain of the Ab respectively comprise the heavy chain and light chain of hRS7.
  35. 根据权利要求1-34中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-34, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中所述Ab包含hRS7。wherein the Ab comprises hRS7.
  36. 根据权利要求1-35中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，The compound of any one of claims 1-35, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof,
    其中所述m通过选自以下组的方法测定：疏水层析、十二烷基硫酸钠聚丙烯酰胺凝胶电泳和液相质谱。wherein the m is determined by a method selected from the group consisting of hydrophobic chromatography, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and liquid phase mass spectrometry.
  37. 一种化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，A compound, or a tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, pro- drug or solvate,
    其中，所述化合物包含选自以下组的结构：wherein the compound comprises a structure selected from the group consisting of:
    Figure PCTCN2022089726-appb-100021
    Figure PCTCN2022089726-appb-100021
    Figure PCTCN2022089726-appb-100022
    Figure PCTCN2022089726-appb-100022
    Figure PCTCN2022089726-appb-100023
    Figure PCTCN2022089726-appb-100023
    Ab为能够结合Trop-2的配体，m为1-8的数。Ab is a ligand capable of binding Trop-2, and m is a number from 1 to 8.
  38. 一种药物组合物，其含有权利要求1-37中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，以及任选地药学上可接受的载体。A pharmaceutical composition comprising the compound of any one of claims 1-37, or a tautomer, meso, racemate, enantiomer, diastereomer, or tautomer thereof The form, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, and optionally a pharmaceutically acceptable carrier.
  39. 含有权利要求1-37中任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐、前药或溶剂合物，和/或权利要求38所述的药物组合物在制备用于治疗和/或预防肿瘤的药物中的用途。Contains the compound of any one of claims 1-37, or a tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof , or a pharmaceutically acceptable salt, prodrug or solvate thereof, and/or the use of the pharmaceutical composition of claim 38 in the preparation of a medicament for treating and/or preventing tumors.
  40. 根据权利要求39所述的用途，所述肿瘤与Trop-2表达相关的肿瘤。The use of claim 39, wherein the tumor is a tumor associated with Trop-2 expression.
  41. 根据权利要求40所述的用途，所述与所述靶点表达相关的肿瘤包含所述靶点高表达的肿瘤和/或所述靶点阳性的肿瘤。The use according to claim 40, wherein the tumor associated with the expression of the target comprises a tumor with high expression of the target and/or a tumor positive for the target.
  42. 根据权利要求39-41中任一项所述的用途，所述肿瘤包含实体肿瘤和/或血液肿瘤。The use of any one of claims 39-41, wherein the tumor comprises a solid tumor and/or a hematological tumor.
  43. 根据权利要求39-42中任一项所述的用途，所述肿瘤选自以下组：肺癌、尿道癌、大肠癌、前列腺癌、卵巢癌、胰腺癌、乳腺癌、膀胱癌、胃癌、胃肠道间质瘤、宫颈癌、食 道癌、鳞状细胞癌、腹膜癌、肝癌、肝细胞癌、结肠癌、直肠癌、结肠直肠癌、子宫内膜癌、子宫癌、唾液腺癌、肾癌、外阴癌、甲状腺癌、阴茎癌、白血病、恶性淋巴瘤、浆细胞瘤、骨髓瘤、或肉瘤。肺癌、尿道癌、大肠癌、前列腺癌、卵巢癌、胰腺癌、乳腺癌、膀胱癌、胃癌、胃肠道间质瘤、宫颈癌、食道癌、鳞状细胞癌、腹膜癌、肝癌、肝细胞癌、结肠癌、直肠癌、结肠直肠癌、子宫内膜癌、子宫癌、唾液腺癌、肾癌、外阴癌、甲状腺癌、阴茎癌、白血病、恶性淋巴瘤、浆细胞瘤、骨髓瘤、和肉瘤。The use according to any one of claims 39-42, wherein the tumor is selected from the group consisting of lung cancer, urethral cancer, colorectal cancer, prostate cancer, ovarian cancer, pancreatic cancer, breast cancer, bladder cancer, stomach cancer, gastrointestinal cancer Stromal tumor, cervical cancer, esophageal cancer, squamous cell carcinoma, peritoneal cancer, liver cancer, hepatocellular carcinoma, colon cancer, rectal cancer, colorectal cancer, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer, vulva cancer, thyroid cancer, penile cancer, leukemia, malignant lymphoma, plasmacytoma, myeloma, or sarcoma. Lung cancer, urethral cancer, colorectal cancer, prostate cancer, ovarian cancer, pancreatic cancer, breast cancer, bladder cancer, stomach cancer, gastrointestinal stromal tumor, cervical cancer, esophageal cancer, squamous cell cancer, peritoneal cancer, liver cancer, hepatocyte cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer, vulvar cancer, thyroid cancer, penile cancer, leukemia, malignant lymphoma, plasmacytoma, myeloma, and sarcoma .
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